Abstract
The results of X-ray structure analysis of metmyoglobin at 300 K, 185 K, 165 K, 115 K and 80 K are reported. The lattice vectorsa andb decrease linearly with temperature whilec shows non-linearity above 180 K, indicating some type of phase transition. Cooling does change the myoglobin structure but only within the structural distribution as determined by individual 〈x 2〉 at room temperature. Two residues showed significant alternative positions for sidechains at higher temperatures while only one position is occupied at low temperatures. In the case of LEU 61 a jump between different positions of the side-chain reduces the potential barrier for the entrance of the O2 molecule to the heme pocket.
The mean square displacements, 〈x 2〉, of the individual residues decrease linearly with temperature in most cases, indicating a parabolic envelope for the potential responsible for motions. A separation of rotational and translational disorder of the entire molecule is discussed. Comparison with Mössbauer spectroscopy indicates that protein dynamics on a time scale faster than 10-7 s is not simply a harmonic process. Extrapolation of the structural distributions toT=0 K shows that a large zero point distribution of the myoglobin structure exists, thus proving that there is no absolute energy minimum for one well defined conformation.
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Ansari A, Berendzen J, Bowne SF, Frauenfelder H, Iben IET, Sauke TB, Shyamsunder E, Young RD (1985) Protein states and protein quakes. Proc Natl Acad Sci USA 82:5000–5004
Artymiuk PJ, Blake CCF, Grace DEP, Oatley SJ, Phillips DC, Sternberg MJE (1979) Crystallographic studies of dynamic properties of lysozyme. Nature (London) 280:563–568
Austin RH, Beeson KW, Eisenstein L, Frauenfelder H, Gunsalus IC (1975) Dynamics of ligand binding to myoglobin. Biochemistry 14:5355–5373
Bartunik HD, Schubert P (1982) Crystal cooling for protein crystallography with synchrotron radiation. J Appl Cryst 15:227–231
Case DA, Karplus M (1979) Dynamics of ligand binding to heme proteins. J Mol Biol 132:343–368
Figgis BN, Reynolds PA, Lehner N (1983) cis-bis(bipyridy)dichloroiron (III) tetrachloroferrate (III), [Fe(bpy)2 Cl2][FeCl4]; structure at 4.2 and at 115 K by neutron diffraction. Acta Cryst B 39:711–717
Frauenfelder H (1985) Ligand binding and protein dynamics. In: Clementi E, Corongiu G, Sarma MH, Sarma RH (eds), Structure and motions: membranes, nucleic acids and proteins. Adenine Press, pp 205–217
Frauenfelder H, Petsko GA, Tsernoglou D (1979) Temperature-dependent X-ray diffraction as a probe of protein structural dynamics. Nature 280:558–563
Frauenfelder H, Hartmann H, Karplus M, Kuntz ID, Kuriyan J, Parak F, Petsko GA, Ringe D, Tilton RF, Connolly MI, Max N (1987) The thermal expansion of a protein. Biochemistry 26:254–261
Hartmann H, Parak F, Steigemann W, Petsko GA, Ringe Ponzi D, Frauenfelder H (1982) Conformational substates in a protein; structure and dynamics of metmyoglobin at 80 K. Proc Natl Acad Sci USA 79:4967–4971
Hartmann H, Steigemann W, Reuscher H, Parak F (1986) Structural disorder in proteins: a comparison of myoglobin and erythrocruorin. Eur Biophys J 14:337–348
Jones TA, Liljas L (1984) Crystallographic refinement of macromolecules having noncrystallographic symmetry. Acta Cryst A 40:50–57
Kendrew JC, Parrish RG (1956) The crystal structure of myoglobin III. Sperm-whale myoglobin. Proc Roy Soc 238A: 305–324
Konnert JH (1976) A restrained parameter structure factor least-squares refinement procedure for large asymmetric units. Acta Crystallogr A 32:614–617
Konnert JH, Hendrickson WA (1980) A restrained thermal factor refinement procedure. Acta Crystallogr A 36:344–349
Krupyanskii Yu F, Parak F, Goldanskii VI, Mössbauer RL, Gaubmann F, Engelmann H, Suzdalev IP (1982) Investigation of large intramolecular movements within metmyoglobin by Rayleigh scattering of Mössbauer radiation (RSMR). Z. Naturforsch 37c:57–62
Levy RM, Sheridan RP, Keepers JW, Dubey GS, Swaminathan S, Karplus M (1985) Molecular dynamics of myoglobin at 298 K. Biophys J 48:509–518
Nienhaus GU, Parak F (1986) Rayleigh scattering of Mössbauer radiation on metmyoglobin. Hyperfine Interactions 29:1451–1454
Nienhaus GU, Drepper F, Parak F, Mössbauer RL, Bade D, Hoppe W (1987) A multiwire proportional counter with spherical drift chamber for protein crystallography with X-rays and gamma-rays. Nucl Instrum Methods A256:581–586
Nyborg J, Wonacott AJ (1977) In: Arnd UW, Wonacott AJ (eds) The rotation method in crystallography. North-Holland, Amsterdam, pp 139–145
Parak F, Knapp EW (1984) A consistent picture of protein dynamics. Proc Natl Acad Sci USA 81:7088–7092
Parak F, Thomanek UF, Bade D, Wintergerst B (1977) The orientation of the electric field gradient tensor in CO-liganded myoglobin. Z Naturforsch 32c:507–512
Parak F, Knapp EW, Kucheida D (1982) Protein dynamics. Mössbauer spectroscopy on deoxymyoglobin crystals. J Mol Biol 161:177–194
Parak F, Fischer M, Graffweg E, Formanek H (1987a) Distributions and fluctuations of protein structures investigated by X-ray analysis and Mössbauer spectroscopy. In: Clementi E, Chin S (eds) Structure and dynamics of nucleic acids, proteins and membranes. Plenum, New York, pp 139–148
Parak F, Hartmann H, Nienhaus GU, Heidemeier J (1987b) Structural fluctuations in myoglobin. In: Ehrenberg A, Rigler R, Gräslund A, Nilsson L (eds) Structure, dynamics and function of biomolecules. Springer, Berlin Heidelberg New York Tokyo, pp 30–33
Phillips SEV (1980) Structure and refinement of oxymyoglobin at 1.6 A resolution. J Mol Biol 142:531–554 Coordinates taken from the Protein Data Bank
Reinisch L, Heidemeier J, Parak F (1985) Determination of the second order Doppler shift of iron in myoglobin by Mössbauer spectroscopy. Eur Biophys J 12:167–172
Ringe D, Petsko GA, Kerr DE, de Montellano PRO (1984) Reaction of myoglobin with phenylhydrazine: a molecular doorstop. Biochemistry 23:2–4
Schwager P, Bartels K (1975) Refinement of setting angles in screenless film methods. J Appl Crystallogr 8:275–280
Schwager P, Bartels K (1977) In: Arnd UW, Wonacott AJ (eds) The rotation method in crystallography. North-Holland, Amsterdam, pp 105–117, 139–151
Steigemann W (1974) Dissertation TU München. Die Entwicklung und Anwendung von Rechenverfahren und Rechenprogrammen zur Strukturanalyse von Proteinen am Beispiel des Trypsin-Trypsininhibitor-Komplexes, des freien Inhibitors und derl-Asparaginase
Swaminathan S, Craven BM, McMullan RK (1984) The crystal structure and molecular thermal motion of urea at 12, 60 and 123 K from neutron diffraction. Acta Crystallor B 40:300–306
Takano R (1977a) Structure of myoglobin refined at 2.0 A resolution. Crystallographic refinement of metmyoglobin from sperm whale. J Mol Biol 110:537–568 Coordinates taken from the Protein Data Bank
Takano R (1977b) Structure of myoglobin refined at 2.0 A resolution. Structure of deoxymyoglobin from sperm whale. J Mol Biol 110:569–584 Coordinates taken from the Protein Data Bank
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Dedicated to Prof. H. Frauenfelder on his 65th birthday
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Parak, F., Hartmann, H., Aumann, K.D. et al. Low temperature X-ray investigation of structural distributions in myoglobin. Eur Biophys J 15, 237–249 (1987). https://doi.org/10.1007/BF00577072
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DOI: https://doi.org/10.1007/BF00577072