Library

Notes: Investigations have been made on the optical rotatory dispersion properties of E. coli ribosomes and their constituent RNA and proteins. The results indicate that (1) no conformational changes are involved in the formation of a 70S particle from the 50S and 308 subunits, (2) the E. coli ribosomal proteins are similar to most globular proteins with little α-helix content, and (3) the conformation of RNA and proteins inside the ribosome is very similar to that in the free state.

Notes: The iodine which is added to an aqueous amylose solution is bound only partly by the amylose while forming the blue complex and partly remains free. The equilibrium normality of the free and the bound iodine at half-saturation of amylose by iodine is designated as [If]v and [Ib]w, respectively. The stability of the poly iodine chain formed within the axis of amylose helices depends on its length, i.e., indirectly on the DP of the amylose helices: the greater this stability, the lower the [If]v value. The amylose molecule consists of helical segments. Such a molecule may behave as a random coil. The average length of the helical segments in freshly prepared amylose-iodine complexes depends on temperature, pH, iodide concentration, the presence of other complex-forming agents, and the DP of the amylose. This latter factor is investigated in the present paper. By the aid of an automatically recording photometrictitrating device the coherent values of [Ib] and [If] were determined. Plotting these values against DPn for mechanochemically degraded as well as for periodateo-xidized amyloses resulted in curves consisting of two linear sections. The break of the curves occurred between DPn 110 and 130. It was concluded that below DPn = 100 the DP of helical segments (= sDPn) is identical to the DPn of the total molecule, i.e., the molecule consists of only a single, relatively stiff helix. Above this limit the molecule contains several helical segments. The DP of these helical segments can be calculated as follows: sDPn = 141.1 - 10.2 × 105[If]v. This equation is considered to be valid for 0.5-0.6 mg. amylose in 100 ml. 0.1N HCl at 20°C., λ = 650 mμ, euuvet diameter 3.4 cm., the feed rate of the iodate-iodide titrating solution (in acid medium resulting in a 5 × 10-3N I2 solution with a molar iodide to iodine ratio of 1.5) is 0.4ml./min. Amylose molecules of, e.g., DPn = 1380 consist of an average of 11.4 segments having a DP of about 120 and consisting of an average of 15-18 helical turns.

Notes: The electrochemical possibilities for investigation of nucleic acids with high molecular weight are restricted to the determination of the adsorption behavior. According to our experience the alternating current polarography (Breyer-polarography) is mainly appropriate for the characterization of changes in the secondary structure of DNA. The Breyer-polarogram shows the alternating current of the dropping electrode in dependence on their potential which varied from 0-2 v. negative against the normal calomel electrode (NCE). By addition of native DNA to the supporting electrolyte (buffer solution) the current drops down in the range of adsorption between 0 and 1 v. At 1.16 v. against NCE the desorption takes place together with the formation of a rounded desorption peak. The investigation was carried out in phosphate buffer solution 0.1m with 0.075m NaCl or in a phosphate buffer 0.18m with 0.03m NaCl. In the pH range above pH 8 NaOH was added to realize the higher pH values. A calf thymus DNA sample having a mean molecular weight of about 18 million was used. The concentration of DNA was 5 × 10-3-1 × 10-1 wt.-%. The polarographic measurements were performed with an a.c./d.c.-polarograph “GWP 564” from Akademiewerkstätten für Forschungsbedarf der Deutschen Akademie der Wissenschaften zu Berlin (DAW). The denaturation of the double helix causes a sharp desorption peak at negative potentials of the alternating current polarogram. This new criterion for the helix-coil transition is due to formation of unpaired bases. These nearly free bases undergo a specific adsorption and the desorption takes place within a narrow potential range. Nevertheless, at present time an electron transfer to particular bases cannot be excluded at special conditions. The increase of the sharp peak permits to estimate: (a) the melting curve of the double helix in agreement with spectroscopic measurements; (b) the photolysis of the double helix; (c) the strand separation in acid and alkaline solution. In the alkaline range the sharp peak increases and reaches its maximum at pH 〉 12. In the acid range, however, no sharp peak is observed and the rounded desorption peak decreases. Therefore, the best way of following the conformation changes is to measure the current difference between the curves of the solutions with and without DNA at electrocapillary-zero-potential. On the classical d.c.-polarogram one can measure small current steps only, which may be caused mainly by capacity changes. Moreover, the scission of the molecule by ultrasonic action can be followed. In this case the rounded peak of DNA increases but the sharp peak does not appear. Similar alternating current polarograms are obtained with poly-A in the native state, because helical and unordered regions coexist in the same molecule. The very rapid indication of these structure changes allows one to carry out kinetic measurements at a fixed potential with this method.

Notes: The theory of Kirkwood for the translational frictional coefficients of structures composed of subunits has been generalized in two ways in order to consider aggregates of nonidentical subunits. One of these generalizations fails when the sizes of subunits are too disparate; the other, derived from a surface shell distribution of frictional elements, is effective over the whole range of relative sizes. It is shown that, in the limit of a continuous surface distribution, a shell model reproduces Stoke's law for a sphere. Comparison is made between the frictional coefficients of spheres, ellipsoids, and rods modeled by finite numbers of subunits and by continuous shells of frictional elements, and those calculated from other theories. Agreement is generally good, though the shell model for prolate ellipsoids of revolution deviates by a few per cent from the Perrin value.

Notes: The method of matrix rank analysis has been applied to a large body of experimental data to determine and identify the minimum number of independent spectral components that, were contained within it. The optical rotatory dispersion of TMV RNA was measured in the wavelength region 230-350 mμ, for a wide range of temperatures and ionic strengths. Over the whole range of conditions considered, the observed optical rotatory dispersion spectra were found to be a superposition of two basic spectra only. Evidence is put forward relating these spectra to the single-strand and double-strand helical forms of the molecule. A model is proposed to explain the observed spectral changes in terms of an equilibrium between these two conformations, which leads to a direct calculation of the percentage composition of the double strand at any of the conditions considered. This equilibrium, while simple in general description, appears multi-state and complex in detail. This method of analysis is both powerful and of wide applicability, since it is independent of the source of the data.

Notes: An infinite series solution to the Mason-Weaver equation is presented for the case in which a synthetic boundary is formed originally between solution and solvent. Digital computations based on this series, and confirmed independently, have been made for a range of parameters. For given conditions, the maximum rate of change of concentration at the meniscus and the time at which it occurs can be easily estimated by means of the curves presented. In equilibrium experiments which commence with formation of a sharp boundary, this enables the fringes to be identified with certainty.

Notes: The course of the reversible helix formation of poly(γ-benzyl L-glutamate) (PBG) dissolved in a mixture of dichloroacetic acid (DCA) and 1,2-dichloroethane (EDC) was followed by measuring the heat capacity and the optical rotation of the system through the transition region. The results of these measurements indicate that the transition enthalpy ΔH the transition temperature Tc, and the Zimm-Bragg parameter σ depend considerably on the PBG concentration as well as on the composition of the solvent. For the standard state of infinite dilution, however, a linear extrapolation of the measured ΔH if values results in a standard value ΔH° = 950 cal./mole, independent of the solvent composition. The results of the calorimetric measurements are discussed in relationship to changes in optical rotation. Some peculiarities in the measured thermodynamic and optical properties in solutions with relatively high content of dichloroacetic acid are reported.

Notes: Helical conformations of infinite polymer chains may be described by the helical parameters, d and θ (the translation along the helix axis and the angle of rotation about the axis per repeat unit), pi (the distance of the ith atom from the axis), dij, and dij (the translation along the axis and the angle of rotation, respectively, on passing from the ith atom to the jth). A general method has been worked out for calculating all those helical parameters from the bond lengths, bond angles, and internal-rotation angles. The positions of the main chain and side chain atoms with respect to the axis may also be calculated. All the equations are applicable to any helical polymer chain and are readily programmed for electronic computers. A method is also presented for calculating the partial derivatives of helical parameters with respect to molecular parameters.

Notes: A dilatometric technique is employed to measure the volume changes occurring on denaturation of collagen solutions and tendons. Partial specific volumes of tropocollagen are calculated to be 0.686 ml./g. in water and 0.689 ml/g in 0.1M citrate buffer. From data on non-aqueous solutions, it is estimated that most of the volume change arises from changes in polymer configuration rather than from changes in solvation.

Notes: We have generated solutions to the Lamm equation to examine the effects of concentration dependence on velocity experiments. Two forms of c dependence are considered: s/s0 = 1 - kc and s/s0 = (1 + kc)-1. Features of these solutions are discussed. The magnitude of the errors resulting from the usual procedure of measuring the rate of movement of schlieren maxima or of the position at which the concentration is one half the plateau value have been examined. These errors are usually negligible after sufficient centrifugation time. The errors in using the half-plateau concentration are less than those using the movement of the peak. We have also examined a technique due to Fujita for determining D from boundary spreading when s/s0 = (1+kc)-1. This method is satisfactory when s/s0 is actually of this form, or under certain limitations when s/s0 = (1 + kc)-1. Creeth has shown that under certain conditions the concentration gradient, curve remains virtually unchanged in shape after separating from the meniscus. When this occurs it is possible to estimate s/D from the data. The condition for such a steady state is that kc0 be sufficiently large. Numerical confirmation of this method is presented in the final section.

Notes: Preliminary experiments have been carried out which measure the variation in the intensity of the light scattered by DNA solutions under the influence of an electric field. Data have been collected on the length of DNA molecules, their electric polarizability, and their dispersion behavior.

Notes: Information concerning the structures of rA-rU, rA-rU2 rI-rC, rA-rI2, and acid rA helices in solutions is reported. Through the use of diquaternary ammonium salts of the general structure, \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_2 {\rm R}_3 \mathop {\rm N}\limits^ + ({\rm CH}_2 )n\mathop {\rm N}\limits^ + {\rm R}_1 {\rm R}_2 {\rm R}_3 \cdot 2{\rm Br}^ - $\end{document} (I), it is shown that (1) the distances between adjacent negatively charged oxygen atoms on the helix increases in the following order rA-rI2 〈 rI-rC 〈 rA-rU ≤ rA-rU2; (2) the density of the helices increases in the order. rA-rI2 〈 rA-rU 〈 rA-rU2 〈 rI-rC; (3) there is a large hydrophobia site in rA-rI2 and possibly also in rA-rU, rA-rU2, and rI-rC helices; (4) the results of the interactions between the salts of type I and the helices may be formulated in semi-quantitative terms by the use of two parameters, α, and β which are shown to be related to the charge separation and the density of the helices, respectively; (5) the studies in solutions compare favorably with the x-ray studies on the fibers; and (6) the acid rA helix differs significantly from the other helices by the fact that the electrostatic interstrand interactions between the negatively charged oxygen atom of a phosphate group and the positively charged 10-amino group of adenine contribute significantly to the stabilization of the helix, and thus it is found that the presence of the salts, I, leads to a significant destabilization of the acid rA helix.

Notes: The half molecules of 186 DNA have been isolated by the Hg(II)-Cs2SO4 density gradient centrifugal ion technique. The buoyant densities of the two halves in CsCI at 25°C. are 1.713 and 1.709 g./cm.3, corresponding to GC contents of 54% and 50%, respectively. Similarly, 5-bromouracil labeled λ DNA halves were separated. The isolation of the four DNA halves made it possible to test for homology in base sequences between the cohesive ends of λ and those of 186. There was no indication of any significant homology in base sequences between the cohesive ends of the two DNA's, as indicated by the absence of a band with intermediate buoyant density in CsCI when either half of 186 DNA was annealed with either half of 5-bromouracil labeled λ DNA and then centrifuged. The lack of cohesion between the two DNA's made it possible to demonstrate unequivocally the formation of interlocked rings (catenanes) between the two DNA's. The existence of a dimeric catenane is evidenced by the formation of a species of intermediate buoyant density when 5-bromouracil labeled λ DNA is cyclized in the presence of cyclic 186 DNA of a relatively high concentration. The molecular weight of one DNA relative to the other can be calculated from the position of the dimeric catenane in a density gradient by using the method of Baldwin. The result was in complete agreement with our previous measurements from the sedimentation coefficients and by electron microscopy. The probability of dimeric catenane formation when one DNA is cyclized in the presence of another DNA is discussed. The experimental results agree with the theoretical expectation.

Notes: Sterically allowed forms of the poly-N-methyl-L-alanine chain were found by calculation of conformational energies as a function of the rotation angles of its chain bonds. The lowest energy form seems to be a right-handed, approximately threefold helix.

Notes: Oriented amylose fibers in the V form were prepared and subjected to x-ray analysis. Unit cells and the probable space group of P212121 were determined for the V anhydrous and V hydrate forms of amylose; the analysis confirms previous predictions of these structures based on x-ray powder patterns. Chain folding in V amyloses is discussed in view of crystallographic evidence and folding experiments conducted with space-filling models. Reported also is evidence for amylose helices having diameters intermediate between 13.0 and 13.7 A.

Notes: Conformational energies of the poly-N-methyl-L-alanine chain have been calculated as a function of the rotational angles of its skeletal bonds, taking into account torsional potentials, van der Waals repulsions, and London attractions. Four pronounced minima in the energy were found; the relative magnitudes of the energies at these minima suggest that the preferred conformation of this chain is either a right-handed, approximately threefold helix, or a slightly distorted, left-handed α-helix. Inclusion of an estimate of the dipolar contribution to the total conformational energy does not significantly affect this conclusion.

Notes: The equilibrium normality [If] of free iodine in amylose-iodine complex formation is a function of the length of the polyiodine chains. This length depends on the DP of helical segments of amylose (sDPn). Values of [If] and of the concentration of the bound iodine [Ib] were determined by the continuous photometric titration with automatic recording. Plots of [Ib] versus [If] give an integral distribution curve. Since the relation between [If] and sDPn is known, the graphic differentiation of the [Ib] versus [lf] curve furnishes the differential distribution curve, representing the mass distribution of the helical segments according to their DP. The peak of this curve is characteristic of the percentage and DP of those helical segments, which occur in the largest amount. On the basis of the differential distribution curve the polymolecularity of the investigated sample may be judged. The titration of amylose samples degraded by various methods gives different distribution curves. Titrating mixtures of samples with widely differing average DP values results in differential curves having more than one maximum.

Notes: The electronic absorption and circular dichroism spectra of the DNA-acridine orange complex have been measured over a range of ionic strength, pH, and DNA phosphate to dye (P/D) ratios. Three circular dichroism bands associated with the long wavelength absorption band of acridine orange are induced on complex formation with DNA. Two of the dichroism bands, due mainly to dimeric dye molecules, are favored by low ionic strength, low pH (3.2), and a low P/D ratio (∼3), while the third, deriving primarily from monomeric dye, is optimum at high ionic strength, neutral pH, and a larger P/D ratio (9). The data suggest that monomeric acridine orange binds to DNA in the form of a left-handed helical array with four dye molecules per turn, while the bound dimer has a skewed sandwich conformation which is itself dissymmetric. The stereochemical relations between the bound monomer dye and the DNA are consistent with a modified intercalation model for the DNA-acridine complex.

Notes: The degree of helicity θ of a series of homologous polypeptides as a function of solvent composition was investigated. The polypeptides studied were: poly-N5-(3-hydroxypropyl)-L-glutamine (PHPG) as well as the corresponding 2-hydroxyethyl and 4-hydoxybutyl derivatives (PHKG and PHBG, respectively). PHPG, which is nonhelical in formic acid, attains helicity on addition of relatively small amounts of formates, formamide, and urea to its solution in formic acid. This demonstrates that the high acidity of pure formic acid is largely responsible for its helix-breaking power-probably through protonation of the peptide bonds. In formic acid-water mixtures all three polymers show a maximum in degree of helicity at a mole fraction of about 0.3 formic acid. This is interpreted as being due to interaction between the two helix-breaking solvents, which results in the formation of an inactive molecular species. It is shown that solvent-induced transitions with helicity maxima are predicted by the Bixon-Lifson treatment when applied to this system.

Notes: Dielectric properties of human and horse hemoglobin were studied at frequencies ranging from 20 kc./sec. to 7 Mc./sec. The relative errors in the measurements were usually less than 10-3 even for mildly conducting solutions (10-3M KCl). The experimental setup allowed us variation and measurement of the degree of oxygenation of the protein and to determine its dielectric parameters. Our main conclusion is that it was not possible to find any variation of the dielectric increment for hemoglobin oxygenation levels of 25, 50, 75, and 100%, approximately. This result is at variance with some previous reports. We cannot give the reason for this discrepancy but discuss some possible explanations. The specific dielectric increment, Δεs/c, of human hemoglobin was shown to be significantly smaller than that of horse hemoglobin (0.28 against 0.32). This physical property is lowered with increasing ionic strength I: Δεs/c = 0.28 and 0.20 for I = 10-4 and 10-3, respectively (human protein).

Notes: An expression is derived for the melting point of a polymer when in equilibrium with a solution in which binding of low molecular weight compounds to the polymer takes place. Allowance is made for the possibility that the crystalline polymer itself is a complex. The argument is a purely thermodynamic one and is based on a consideration of the change in free energy as a result of a change in binding. Allowance is made also for non-specific polymer-solvent interactions, in which the mixture of low molecular weight solvents is treated as a single solvent. Special attention is paid to “inverted” melting transitions, i.e., cases in which the melting point increases with increasing dilution of the polymer. It is shown that as a rule this is accompanied by a corresponding, inverted effect of the solvent composition on the melting point. It is further shown that-in the absence of binding, “normal” behavior at the critical point (i.e., phase separation is induced by lowering the temperature) is always accompanied by “normal” melting behavior (i.e., a decrease in melting point when the polymer is diluted). Also, “inverted” melting always implies that phase separation at the critical point is induced by heating, but the reverse is not necessarily true.

Notes: The helix-coil transition temperature Tc of poly(γ-benzyl L-glutamate) in binary solvent mixtures of dichloroacetic acid and 1,4-dichlorobutane, 1-chlorooctane, or 1-chlorododecane have been measured. A treatment is presented with which the transition enthalpy can be calculated from the observed dependence of Tc on solvent composition. Results are compared with previously obtained calorimetric data. The underlying assumptions of the calculation are discussed.

Notes: A marked molecular weight fractionation accompanies the demixing or phase separation resulting from the complex coacervation of mixtures of aqueous solutions of salt-free isoionic unfractionated gelatins with pI's of 5 and 9. Viscosity studies show that the fractionation is such that the concentrated phase tends to maintain constant, homogeneous composition. A second feature, seen in phase volume and concentration measurements, is a marked self-suppression of coacervation intensity with increasing mixing concentration. These data were interpreted in terms of a dilute-phase aggregate model which assumes nearly equal electrostatic free energies of mixing in dilute aggregate and concentrated random phase. The driving force for phase separation is the entropy increase upon formation of the random phase but demixing also depends upon the polymer-solvent interaction parameter χ, in the same fashion as in simple coacervation. The dilute-phase aggregate model indicates that the sharp molecular weight selection takes place in the aggregate formation step and explains the self-suppression. Phase equilibria studies utilizing fractionated, paucidisperse high molecular weight gelatins, emphasize the requirement for concentrated phase homogeneity and indicate that aggregates of different molecular weight may act as different components, so that χP1Q1,PkQk 〉 0, bringing about a separation of the system into three or more coexisting phases. The formation of several coexisting phases from a homologous polyelectrolyte system and the very marked requirement for phase homogeneity suggest that the phenomenon of complex coacervation is a very good model for some of the essential steps in the pre-biologic organization of polymeric polyions.

Notes: Light scattering of multichain poly-α-amino acids was studied in dimethylformamide (DMF). The polymers consisted of a backbone of poly-L-lysine of degree of polymerization n with side chains of benzyl L-glutamate and benzyl L-aspartate of degree of polymerization, m, on each ε-amino group. The backbone length n is known and m is obtained by amino acid analysis. The results on a series of such materials confirm this structure and show that the molecules are dissolved in highly compacted conformations. It was found that DMF is a poor solvent for these polymers. In the case of the higher molecular weight polymers, the solutions initially were not molecularly disperse. The aggregates were resistant to dilution in the experimental range. Mild heat treatment, however, disaggregated the solutions irreversibly, and the light-scattering data indicated that a structural rearrangement of the molecules had occurred.

Notes: The intrinsic viscosity, sedimentation and diffusion of a series of branched, multichain poly-α-amino acids having a poly(L-lysine) backbone and poly(γ-benzyl L-glutamate) and poly (β-benzyl L-aspartate) side chains was studied at room temperature in dimethylformamide. The molecules were found to be extremely compact structures in which the molecular backbone is either lying along the major axis in a slightly twisted configuration (the longer the side chain the smaller the twist) or is coiled up in the form of a disk with backbone and side chains coplanar. Heat treatment (to 70°C.) introduces only small changes in the hydrodynamic parameters showing that the heat-labile aggregates detected by light scattering are reversibly broken up during the hydrodynamic measurements. The above structural information concerns the initial metastable conformation of the molecules which is irreversibly destroyed by heat treatment.

Notes: The optical rotatory dispersion and ultraviolet absorption spectra of the 64 trinucleoside diphosphates containing the bases A, U, C, and G have been calculated by using a simple semiempirical approach. These calculations accurately predict the optical properties of the nine trimers for which extensive experimental results are available. The computed optical data should be useful in the identification of oligonucleotides obtained in the course of sequence determination of ribonucleic acids and should simplify the determination of the concentration of oligonucleotides in aqueous solution. Additional calculations indicate that it should be possible to analyze most, mixtures of sequence isomers of trinucleoside diphosphates by direct, measurement of the ORD of the mixture at neutral pH.

Notes: The equilibrium binding constants of Cd++ and Cu++ to native and denatured calf thymus DNA were determined polarographically. The binding constants are an exponential function of the potential at the binding site and as such they vary with ionic strength and with the charge on the DNA molecule. The correlation between the fraction of sites occupied by heavy metal ions and between the thermal stability of DNA in solution is discussed.

Notes: Complex shear viscosity η* = η′ - iη″ of poly (L-glutarmic acid) solution was measured by the torsional crystal method at 50 kc./sec. as a function of pH. A sharp peak was found at the midpoint of the helix-coil transition region in both η′ and η″. The relaxation time is calculated from η′ and η″ assuming a single relaxation process and the peak value at the midpoint of transition is estimated at 10-6 sec. Such behavior agrees well with the prediction from the theory of Schwarz. The attenuation of longitudinal sound waves at,3 Me./sec. was measured as a function of pH for solutions of poly(glutamic acid), glutamic acid, and gelatin. A small attenuation peak was observed for the three solutions, the peak height being almost, the same for them. The peak is interpreted in terms of the dissociation reaction of side chains.

Notes: We report the circular dichroism (CD) spectrum of poly-Lacetoxyproline in trifluoroethanol, a solvent in which the form I to form II conformational isomerization occurs slowly enough to permit observation of the spectral changes. A comparison is made to poly-Lproline. As judged by the similarity of the CD spectra, the conformations of the corresponding forms of the two polymers must be nearly the same. Transition assignments are proposed; these are shown to agree with the theoretical calculations of Pysh. There is a serious unexplained discrepancy between our solution data and those of Fasman for poly-Lacetoxyproline.

Notes: The dielectric relaxation of DNA solutions has been investigated with and without extraneous ions covering a wide frequency range. The effect of monovalent ions such as Na, K, and Li as well as divalent ions such as Mg, Ca, and Hg have been included in the study. These ions are found to have a profound effect on the dielectric increment and the relaxation time without affecting the molecular dimension drastically. This dielectric effect is interpreted as indicating the importance of counterion fluctuation on the low frequency dielectric constant of DNA in solution. The effect of an organic ion, tetra-methylammoniun bromide, has also been studied. This ion has no noticeable effect. A simple theory is derived on the basis of a microscopic model to account for the effect of external ions on the dielectric behavior of solutions of DNA.

Notes: Block and random copolymers of γ-benzyl L-glutamate and glycine were studied by optical rotatory dispersion methods in order to ascertain the extent and stability of the α-helix formed by these polymers in appropriate solvent mixtures. Results indicate that when in solution such polymers do contain sections of α-helix. However, it appears that glycine does not readily fit into the α-helix which is largely due to the γ-benzyl L-glutamate segments. Further, there are indications that the helical sections in the random copolymers are due to the nonrandomness of the polymerization.

Notes: It is demonstrated that, poly(A + U) and poly(I + C) are both formed under low ionic strength conditions. Continuous variation studies indicate the formation of copper(II) complexes of poly A, poly C, and poly I, but not of poly U. Copper(II) in a 1:1 ratio to polynucleotide prevents the formation of poly(A + U) and brings about the dissociation of the poly (A + U) complex produced in the absence of the metal. Poly (I + C) is similarly dissociated by copper(II) ions. The addition of sufficient electrolyte reverses the copper(II) induced dissociation of poly(I + C). The effect of copper(II) on ordered synthetic polynucleotides is thus very similar to its effect on DNA.

Notes: The synthesis and some of the physical-chemical properties of tricopolymers of L-glutamic acid, L-lysine, and L-alanine are reported here. The molar ratios of the glutamyl: lysyl: alanyl residues were 1:1:X or 3:2:X, where the alanyl content X was increased in regular steps. The α-helix content calculated from the optical rotatory dispersion of the polypeptides is compared with a predicted helix content estimated from the composition of the polymers and the known behavior of the homopolypeptides at pH 3, 8, and 12. At pH 3 copolypeptides containing 20 mole-% or more alanine exhibit a helix content equal to the sum of their alanyl and glutamyl residue contents. At pH 8 the helix content equals the alanyl content when the latter was 40 mole-% or higher; at lower alanyl contents the electrostatic interaction between charged glutamyl and lysyl residues makes some contribution. At pH 12 the amount of helix observed is proportional to the mole ratio of alanine residues present in the polymer. The helix content of a tricopolymer containing 1:1:3 mole ratios of glutamyl: lysyl: alanyl residues was determined in solutions of lithium bromide and in urea solutions. Both reagents led to a decrease in helix content at pH 3 and 8 to a minimum of approximately 20% helix in 8M urea or 5.5M LiBr. The helix-random chain transition curves at pH 3 and 8 are parallel when the urea concentration is varied, but differ in shape when the lithium bromide concentration is varied at pH 3 and 8. The mode of action of these two “denaturing” reagents may thus be different. Heating the same tricopolypeptide at pH 3 or 8 from 5 to 80°C. also led to a helix-random chain transition centered at approximately 45°C.

Notes: Potentiometric titration curves of the silver(I) complexes of cytidine, adenosine, and uridine show little uptake of base below pH 7, unlike the curve for silver(I)-guanosine, which shows extensive base uptake at neutral pH. This observation is correlated with spectrophotometric data showing little difference between the silver complex spectra of adenosine, cytidine, and uridine and the spectra of uncomplexed nucleosides, except at high pH, but showing a great difference between the silver complex spectra of guanosine and inosine and the corresponding uncomplexed nucleosides even at pH 6. Similar comparisons of the silver complexes of poly A, poly C, poly I, and poly U, both by potentiometric titration and by spectrophotometry, show that poly I behaves like guanosine and inosine as expected, differing from poly A and poly C. However, poly U behaves like poly I and thus does not resemble uridine in its complexing behavior. There is thus a dichotomy between poly A and poly C on the one hand in silver complexing phenomena, compared with poly U and poly I on the other. When silver(I) is added to systems containing zinc(II) and various polynucleotides, the same dichotomy is noted. Silver(I) inhibits the degradation by zinc(II) of all four polynucleotides, but the degradation of poly I and poly U is prevented virtually completely. Silver(I) alone has no degradative effect on RNA and inhibits, the zinc(II) degradation of RNA. Polynucleotide complexes in which silver(I) and zinc(II) are simultaneously bound to different positions on the macromolecules are postulated as intermediates in the inhibited degradation reactions.

Notes: C-[2.4.6-Trimethoxy-phenyl]-N-[2.4.6-trinitro-phenyl]-nitrilimin (13) isomerisiert rasch zum 4.6-Dinitro-1-[2.4.6-trimethoxy-benzoyloxy]-benztriazol (22).

Notes: Dimeres Rhodiumdicarbonylchlorid und polymers Iridiumtricarbonylchlorid reagieren mit Kaliumdiphenylphosphid unter KCI-Abspaltung und Bildung von Carbonylkomplexen mit μ-Phosphido-Brücken. [(CO)2RhPR2]n und [(OC)3IrPR2]2 (R = C6H5). Die Einwirkung von Tetraphenyldiphosphin und -diarsin, R2EER2(E = p, As), auf [Rh(CO)2CI]2 führt über instabile Komplexe CI(OC)Rh(E2R4)2 zu dimeren Verbindungen vom Typ [CI(OC)RhER2]2, von denen die cis-trans-Isomeren isoliert werden können. Auch die entsprechenden Iridiumverbindungen wurden über den Pyridinkomplex CI(OC)2Irpyr erhalten.Alkylendiphosphine liefern mit [Rh(CO)2CI]2und CI(OC)2Ir pyr zunächst ohne CO-Abspaltung zweikernige, instabile Komplexe vom Typ CI(OC)2MeR2P - [CH2]N - PR2Me(CO)2CI(Me = Rh, IR; R = C6H5;n = 1 und 2). Mit einem überschuß der Phosphine entstehen die Neutralkomplexe CI(OC)Rh(R2P)2C2H4 und [CI(OC)Rh(R2P)2 CH2]n sowie die kationischen Komplexe [Rh {R2P)2C2H4)}2]X und [OCIr{(R2P)2CH2}2]X (X = Cl, PF6).

Notes: 3.4-Dihydroxy-phenäthylamin reagiert im Gegensatz zu anderen primären aliphatischen Aminen mit Acetessigaldehyd nicht nur zum entsprechenden 2-Methyl-5-acetyl-pyridiniumsalz (1), sondern außerdem unter dehydrierender Verknüpfung von Benzol- und Pyridinring zu einem 6.7-Dihydro-benzo[a]chinoliziniumsalz (2), dessen Methyl- und Acetyl-Seitenkette sich in „natürlicher“ Stellung befinden. Der vollaromatische Ring C von 2 läßt sich mit Natriumborhydrid hydrieren (8, 11), Ring B mit Tetrachlor-o-benzochinon dehydrieren (13, 14), die Acetyl-Seitenkette in eine Äthylgruppe umwandeln (9, 10); mit Allylbromid werden insgesamt vier Allylreste eingeführt. - Fragen des Reaktionsverlaufs, der Spektren sowie der Verwandtschaft zu Alkaloiden werden erörtert.

Notes: Der Verlauf massenspektrometrischer Abbaureaktionen ist von der durchschnittlichen Angregungsenergie der Molekül-Ionen abhängig. Diese setzt sich aus der vom angreifenden Elektron auf das Molekül übertragenen, das Ionisierungspotential überschreitenden Energiemenge, und der thermischen Anregung der Moleküle vor der Ionisation zusammen. Durch Verminderung dieser Anregungsenergie können mehrstufige massenspektrometrische Abbauprozesse auf Zwischenstufen abgefangen werden, wodurch ein besserer Einblick in den Zerfallsmechanismus gewährleistet wird. Grundlagen und Anwendungsbereich dieser Methode werden am Beispiel aliphatischer Äther erläutert.

Notes: Ausgehend von Acetylencarbonylverbindungen werden einige disubstituierte Thiophen-Derivate (1-3) dargestellt. Weiterhin wird die Reaktion von Polyinen mit Natriumdisulfid untersucht. Als Hauptprodukte erhält man Dithiacyclopenten-Derivate (8, 15-17). Die Eigenschaften dieser Substanzklasse werden diskutiert.

Notes: Die Synthese linearer Furanoxanthone (5-Oxo-5H-furo[3.2-b]xanthene) aus 2-Hydroxy-3-acetyl-xanthonen wird an den Beispielen 5-Oxo-3-methyl- und 5-Oxo-3.7-dimethyl-5Hfuro[3.2-b]xanthen (4a, b) beschrieben.

Notes: Die IR-Spektren m- und p-substituierter Phenylessig- und Phenylglyoxylsäuren werden im Bereich der Carbonyl-und Hydroxylbanden verglichen. Die streng proportionalen Frequenzverschiebungen von α-Carbonyl- und Hydroxylbanden im Falle der chelatisierten Phenyl-glyoxylsäuren zeigen, daß die Wasserstoffbrücke zum π-Elektronenpaar der Carbonylgruppe gebildet wird, und daß sowohl induktive wie konjugative Effekte über intramolekulare Wasserstoffbrücken mit hoher Empfindlichkeit vom Protonenacceptor zum -donator übertragen werden.

Notes: Die stellungs- und konfigurationsisomeren n-Octene wurden mit konz. Schwefelsäure sulfatiert und die Isomerenverteilung sowohl in den sich bildenden Alkylsulfaten als auch in dem nicht umgesetzten Olefin bestimmt. Läßt man das Olefin mit einem Unterschuß oder höchstens mit der äquimolaren Menge an Schwefelsäure so reagieren, daß die Schwefelsäure dem Olefin zugefügt wird, bilden sich vornehmlich die aufgrund der Doppelbindungslage im eingesetzten Olefin zu erwartenden Alkylsulfat-Isomeren. Wird hingegen das Olefin der Säure zugefügt bzw. ist diese am Ende im Überschuß vorhanden, so bildet sich unabhängig von der Doppelbindungslage und der Konfiguration des Ausgangsolefins stets das gleiche Isomerengemisch sekundärer Alkylsulfate. Das Ausmaß der Bindungsisomerisierung im nicht umgesetzten Olefin entspricht etwa der Isomerenverteilung im Sulfatestergemisch.

Notes: Die Synthese der Tetrazolylmercapto-essigsäure-azide 5a-c und Tetrazolylmercaptomethyl-isocyanate 6a-c wird beschrieben.

Notes: Durch Reaktion von (CH3)3SiN3 mit SnCl4 erhält man über eine unter bestimmten Bedingungen isolierbare Zwischenverbindung Cl3SnN3 (CH3)3SiN3 als Endprodukt [Cl2Sn(N3)2]n. Einige Eigenschaften und das Infrarotspektrum der Verbindungen sowie Strukturvorschläge werden beschrieben.

Notes: Die Dehydrierung von Phorbol und Phorbol-f-ol sowie die Dehydratisierung von Phorbol-f-ol führt zu Azulenen. Dies deutet darauf hin, daß Phorbol möglicherweise eine Hydroazulen-Struktur enthält.

Notes: 1.3-Dioxolan (1a) und 2-Alkyl-1.3-dioxolane (1b-f) sind sehr leicht peroxygenierbar. Als Primärprodukte der Peroxygenierung wurden die Hydroperoxide (2a-f) isoliert. Diese gehen durch Hydrierung quantitativ in Glykolhalbester über. Unter den Bedingungen der Vakuumdestillation bilden sich die dimeren Peroxide (5) und H2O2.

Notes: Die bisher untersuchten Arten des Tribus Helenieae zeigen, daß die botanische Unterteilung dieses Tribus gut mit den gefundenen Inhaltsstoffen übereinstimmt. Neben bereits bekannten Polyinen werden einige neue Verbindungen (1,2,6,10) isoliert, die sich biogenetisch von dem weitverbreiteten Pentainen 3 ableiten.

Notes: Die Umsetzung von Dehydrochinolizidin (2) mit dem Hexadienon 1 liefert ein tricyclisches Keton (3), das in der Enolform 4 vorliegt und äußerst leicht in ein Diketon (10) übergeht.

Notes: In den 2-Thio-hydantoin-Derivaten 2 von Asparagin-, Glutamin- und α-Amino-adipinsäure kann das Ringsystem als kombinierte Schutzgruppe für die α-Amino- und die benachbarte Carboxyl-Funktion betrachtet werden. Am noch freien Carboxyl können die Verbindungen 2 mit Aminosäureestern Peptidverknüpfungen eingehen. Dies kann auf konventionelle Weise erfolgen, während beim Glutamin- und α-Amino-adipinsäure-Derivat die zugehörigen Lactame 3 als ω-carboxylaktivierte Formen fungieren können. Die Entschwefelung der Thio-hydantoin-ω-peptide 5 mit Raney-Nickel liefert ω-Peptidester, die in Form der α-Formamino-carbonamide 4 anfallen. - Ungesättigte, methylverzweigte 2-Thio-hydantoin-peptidester 8 lassen sich analog aus den Kondensationsprodukten 7 von 2-Thio-hydantoin mit Brenz-traubensäure, Acetessigsäure-äthylester oder Lävulinsäure gewinnen. Die Entschwefelung bietet eine Möglichkeit zur Synthese β-methylverzweigter Analoger der erwähnten ω-Peptidester (Beispiel 8b). - Aus optisch aktiver Pyroglutaminsäure und N-Benzoyl-L-alanin können zwar aktive Thiohydantoin-Derivate dargestellt werden, jedoch racemisieren diese leicht. - Die Entschwefelung von 4-Cyclohexylimino-5.5-dimethyl-2-thio-hydantoin (14, R = R′ = CH3, R″ = c-C6H11) liefert 4-Cyclohexylimino-5.5-dimethyl-4.5-dihydro-imid-azol (17).

Notes: Die Strukturen und Konfigurationen einiger natürlicher Schwefelverbindungen (2, 3, 4, 26 bis 28) werden durch Synthese gesichert.

Notes: Die Wurzeln von Serratula radiata Bieb. enthalten neben dem weitverbreiteten En-tetrain-en 1, mehrere Thiophenverbindungen, von denen drei (5, 7, 8) bisher noch nicht bekannt waren. Die Strukturen werden durch die physikalischen Daten sowie durch einige Reaktionen geklärt.

Notes: Acetessigsäure-äthylester wird an der Methylengruppe leicht nitrosiert. Auf die Explosionsgefahr bei der Reinigung des Reaktionsproduktes durch Hochvakuumdestillation wird jedoch nirgendwo hingewiesen. Am tert.-Butylester wird der Zerfallsmechanismus untersucht.

Notes: Nach weiteren Untersuchungen über Derivate, besonders aktivierter Ester, und Peptide von NHPhe wurde ausgehend vom C-terminalen Tetrapeptid 5 mit Hilfe des Boc-NHPhe-hydroxysuccinimidesters (2c) das [5-Asparagin, 7-L-α-Hydrazino-β-phenyl-propionsäure]Eledoisin-(4-11)-Octapeptid (als Bis-trifluoracetat9b) nach der Nitrophenylester-Methode aufgebaut.

Notes: Die Addition von Dichlormethyllithium (20) und verschiedener olefinischer Carbenoide (8,24,30) an Triphenylbor führt zu α-chlorierten Lithium-boraten, die unter „inverser Stevens-Umlagerung“ zu Boranen und Lithiumchlorid thermolysiert werden. Die Substitution des α-Halogens durch Bor-ständiges Phenyl verläuft stereospezifisch unter Konfigurationsumkehrung und wird als Synchronprozeß interpretiert. Ein zweites Halogen in α-oder β-Stellung veranlaßt Folgereaktionen am umgelagerten Boran.

Notes: Die magnetische Protonenresonanz der substituierten Diaziridine 2-81) zeigt, daß der gesättigte C—N—N-Dreiring ein Asymmetriezentrum darstellt. Die Inversion an mindestens einem der beiden Stickstoffatome verläuft demnach verhältnismäßig langsam. Die freie Enthalpie der Aktivierung muß bei einigen der untersuchten Verbindungen größer als 21 kcal/ Mol sein.

Notes: Die chemischen Verschiebungen der Ringprotonen von 36 substituierten 1.3-Diphenyl-Δ2-pyrazolinen und von 23 substituierten 3-Phenyl-Δ2-isoxazolinen werden mit denen der Stammkörper verglichen. Die Methode der kleinsten Fehlerquadrate führt zu den Parametern Δτ für gem., cis-vic. und trans-vic. Substituenten. Diese Substituenteneffekte werden theoretisch diskutiert und sind von diagnostischem Wert bei der Strukturermittlung.

Notes: Durch Addition von Vinyldiazomethan an Chinone werden 3-Vinyl-pyrazolochinone (1-4) dargestellt. Ihr chemisches Verhalten wird untersucht.

Notes: Dreizehn neue Verbindungen des Typs 4-F—C6H4—X, mit X = PH2, P(CH3)2, P(C6H5)2, P(O)(C6H5)2, P(O)(CH3)2, P(O)(OC2H5)2, P(S)Cl2, P(S)(C6H5)2, P(CH6)3J, P(C6H4F)2, P(O)(C6H4F)2, P(S)(C6H4F)2 und P(CH3)(C6H6H4F)2J wurden dargestellt. Von den genügend löslichen Substanzen sowie von 4-F—C6H4—PCI2 und 4-F—C6H4—P(O)Cl2 wurden die 19F-NMR-Spektren aufgenommen, die chemischen Verschiebungen, bezogen auf Fluor-benzol, bestimmt und daraus die Hammettschen σp-Konstanten der phosphorhaltigen Substituenten berechnet. Damit war zum ersten Mal ein quantitativer Vergleich der PH2-mit der NH2-Gruppe möglich. Alle diese Substituenten sind Elektronenakzeptoren, der stärkste gemessene Akzeptor ist die P(O)Cl2- und der schwächste die P(CH3)2-Gruppe.

Notes: Perjodatoxydation von 3-[β-D-Glucopyranosyl]-thymin und anschließende Nitromethan-Cyclisierung führt zu einem Gemisch von 3-[3-Nitro-3-desoxy-hexopyranosyl]-thyminen, aus dem durch Schichtchromatographie das gluco-Isomere 3 abgetrennt wurde. Katalytische Hydrierung liefert 3-[3-Amino-3-desoxy-β-D-glucopyransoyl]-thymin (5). Die Konfigurations-zuordnung ergab sich aus NMR-spektroskopischen Daten des Tri- und Tetraacetats (6 bzw. 4).

Notes: Ausgehend von 7-[β-D-Ribofuranosyl]- (1) bzw. 7-[β-D-Glucopyranosyl]-theophyllin (3) wurde durch Perjodatoxydation und anschließende Nitromethan-Cyclisierung ein Gemisch von 7-[3-Nitro-3-desoxy-hexopyranosyl]-theophyllin-Isomeren erhalten. Durch fraktionierte Kristallisation ließen sich die gluco-, manno- und galakto-Verbindungen (6, 4, 5) trennen. Katalytische Hydrierung führte zu den entsprechenden [3-Amino-3-desoxy-β-D-hexopyranosyl]-theophyllin-Nucleosiden (9, 7, 8), die als Hydroacetate, N-Acetate und Tetraacetate charakterisiert wurden. Die Konfigurationszuordnung ergab sich aus vergleichender Papierchromatographie, NMR-spektroskopischen Daten und chemischen Identifizierungsreaktionen.

Notes: Durch Alkali-Einwirkung auf 2.4.6-Triphenyl-o-chinolacetat (3) in Gegenwart von Luftsauerstoff entsteht ein farbloses Dimeres des 3.5-Diphenyl-2-benzoyl-cyclopentadienons (8) der Konstitution 9a oder 9b. Dieses steht in Lösung mit seinem roten Monomeren in einem temperaturabhängigen Gleichgewicht, zeigt also Thermochromie. Unter Sauerstoffausschluß bildet sich aus o-Chinolacetat 3 das entsprechende vollständig enolisierte Diphenyl-benzoyl-cyclopentenon 5. Reaktionen, IR- und NMR-Spektren werden beschrieben.

Notes: Die Darstellung von 1.1-Dihalogen-cyclopropanen aus Olefinen, Trihalogenmethan und Äthylenoxid in Gegenwart eines salzartigen Katalysators wird beschrieben. Die Reaktionen werden über intermediär gebildetes Dichlor-, Fluorchlor- und Difluorcarben formuliert.

Notes: Eine modifizierte Friedel-Crafts-Acylierung am Δ3-Caren ergibt in hohen Ausbeuten 3-Acyl-Δ4-carene. Diese Verbindungen ermöglichen die Synthese zahlreicher 3-substituierter Derivate des Δ3-Carens, des Carans und des Δ4-Carens.

Notes: S.S'-Dialkyl-dithiobiurete (1), durch Alkylierung von Dithiobiuret gut zugänglich, eignen sich als Ausgangsstoffe für eine einfache Synthese von 1.3.5-Triazinen, die mehrere funktionelle Gruppen enthalten. Mit Carbonsäurechloriden lassen sich in einem Reaktionsschritt 2-substituierte 4.6-Bis-alkylmercapto-1.3.5-triazine (2, 4) darstellen, die mit Hydroperoxid in die 4.6-Dihydroxy- (12) und mit Chlor in die 4.6-Dichlor-1.3.5-triazine (13) übergeführt werden können. Die Anwendung der neuen Synthese auf S-Alkyl-guanyl-isothiuronium-salze (8) und Imino-bis-[carbimidsäure-phenylester] (10) führt zu 2-substituierten 4-Amino-6-alkylmercapto-(9) bzw. 4.6-Diphenoxy-1.3.5-triazinen (11).

Notes: 4-Chlor-5-cyan-pyrimidin (3) gibt mit Ammoniak, prim. und sek. Aminen, Hydrazinen, Natriumalkoholaten sowie Natriummercaptiden unter Substitution des 4-Chlor-Atoms die Derivate 4-14. Durch katalytische Reduktion des 4-Amino- und 4-Hydroxy-5-cyan-pyrimidins erhält man den bisher nicht bekannten 4-Amino- und 4-Hydroxy-pyrimidin-aldehyd-(5) (18 und 17).

Notes: Die Produkte der Addition von Tetramethylthioharnstoff an Acetylendicarbonsäure-di-methylester in Dioxan als Solvens werden beschrieben, die Konstitutionen aufgeklärt und ihre Bildung diskutiert.

Notes: Die Jod-Oxydation von Tetramethoxy-äthylen (1) ergab Dimethyloxalat (4) und Tetramethoxy-bernsteinsäure-dimethylester (6). Durch Oxydation in Gegenwart von Natriummethylat ließ sich dabei die Zwischenstufe des Tetramethoxy-äthylen-Dikations 3 zu Hexamethoxyäthan (7) abfangen. Die Reduktion weiterer Verbindungen mittels Tetramethoxy-äthylen wurde untersucht.

Notes: An Perchlorallen (1) lassen sich im Temperaturbereich von -75 bis -30° Brom und Chlorjod sowie in Gegenwart von AICI3 Chlorwasserstoff und aliphatische Säurechloride addieren, ohne daß die Dimerisation zu 2 merklich in Erscheinung tritt. Die Additionsrichtung der unsymmetrischen elektrophilen Agenzien, die geklärt wird, deutet auf eine hohe Elektronendichte am C-2 des Allens hin. Im Zuge der Konstitutionsaufklärung des aus Acetylchlorid und 1 gebildeten Methylketons 8a werden einige hochchlorierte Ketone (9) und Carbonsäuren (11, 12, 13) beschrieben. Das Allendimere 2 reagiert mit Diazomethan bei 0° unter N2-Abspaltung zu dem Spiran 17 mit Cyclopropanring.