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Notes: Generalized fatigue damage is derived based on S-N curve under constant amplitude loadswith S characterizes the load cycle by the combination of load peak and load amplitude. For randomfatigue loads, a natural choice is to use half cycle (Range) counting method. Rain Flow counting ispopular due to its useful nature to pick-out full cycles among random loads. It is found in the presentresearch through analysis and verified by experimental results that Rain Flow counting seriouslyoverestimate the damage for the actual true random loads owing to its inherent nature and the Rangecounting method is more reliable. As the filtering threshold increases gradually, the overestimationeffect by Rain Flow counting method becomes less, and the calculated damages by Rain Flow andRange counting begin to converge. The filtering threshold damage corresponding to the “fatiguelimits” for the concerned case was calculated. According to equal damage principle, the filteringthreshold amplitudes were derived and found to vary with the load peaks. In practice, an acceptablesolution is to use quasi-variable filtering thresholds

Notes: Crystals of a soluble monomeric quinocytochrome alcohol dehydrogenase (ADH-IIB) and of a trimeric membrane-associated quinocytochrome alcohol dehydrogenase (ADH-GS) have been obtained. The ADH-IIB crystals are triclinic, with one monomer in the unit cell, and were obtained in the presence of PEG 8000, sodium citrate, HEPES buffer and 2-propanol. X-ray data were collected at 110 K to 1.9 Å resolution (Rmerge = 6.4%) and the orientation of a methanol dehydrogenase search molecule (from Methylophilus methylotrophus W3A1) was obtained by molecular replacement. Preliminary refinement of this model (10.0–3.0 Å resolution, R = 0.37, Rfree = 0.40) led to tentative identification of the two highest peaks in a native anomalous difference Fourier map as the Fe atom of the heme and a calcium ion interacting with the PQQ prosthetic group. The ADH-GS crystals are tetragonal, displaying six similar lattices, both primitive and centered, and were grown by the sitting-drop method after replacement of Triton X-100 by dodecylmaltoside or octaethylene glycol monododecyl ether in the presence of ammonium sulfate and sodium acetate buffer, with and without PEG 3500 and calcium ion. The best diffraction is obtained at 110 K where the resolution extends to about 4 Å in the a and b directions and about 3 Å in the c direction.

Notes: Azurin from Pseudomonas putida is a blue copper protein which functions as an electron carrier. Two crystal forms of azurin were grown, one in the presence and the other in the absence of zinc acetate; each belongs to space group P21 and contains two molecules per asymmetric unit. The zinc-free crystals have cell dimensions a = 43.25, b = 50.65, c = 54.60 Å, β = 107.79°, while the crystals grown from zinc-containing solution have cell dimensions a = 40.76, b = 51.22, c = 54.96 Å, β = 103.12°. The latter crystals were found to have four zinc ions incorporated into the crystal lattice. Both crystal structures were solved by the molecular-replacement method using the program MERLOT. The search model was the structure of azurin from Alcaligenes denitrificans. The crystallographic R factor for native azurin is 0.169 (Rfree = 0.257) from 8 to 1.92 Å resolution, while that for zinc azurin is 0.181 (Rfree = 0.248) from 10 to 1.6 Å resolution; for each structure the root-mean-square deviation in bond lengths from ideal values is 0.007 Å. In both crystal structures the Cu atom forms three strong bonds in the equatorial plane, two with N^{\delta1} from His46 and His117, and one with the thiolate S atom of Cys112. Two longer axial approaches are made by the S^\gamma from Met121 and the carbonyl O atom from Gly45. This results in a distorted trigonal bipyramidal co-ordination around the Cu atom. It further confirms the presence of a weak fifth bond to the copper in P. putida azurin, as with other azurin structures described at high resolution. The N^{\delta1} atom of His35 is protonated, as it is in the low-pH form of azurin from Pseudomonas aeruginosa but unlike the low-pH form of the azurins from Alcaligenes denitrificans or Alcaligenes xylosoxidans. In each crystal form the two molecules of azurin in the asymmetric unit are related by a local twofold axis and form a dimer stabilized by the interaction of a pair of hydrophobic patches surrounding the partially exposed His117 side chain. In the other known azurin crystal structures, analogous dimer formation is observed, but with different relative orientations of the molecules. The four zinc ions introduced during crystallization of zinc azurin are bound to the protein and participate in five- and sixfold ligand coordination with no affect on the copper binding site. The zinc ligands are N^\delta from His, carboxylate O atoms from Asp and Glu, O^\gamma from Ser and water molecules. One of the zinc ions, located on a non-crystallographic twofold axis, links the dimers of the asymmetric unit into continuous chains parallel to the crystallographic (−101) direction and is primarily responsible for the altered unit-cell parameters. Two of the other zinc ions bind to His83, one in each molecule.

Notes: [Auszug] The X-ray structure of the homotetrameric lysosomal acid hydrolase, human β-glucuronidase (332,000 Mr), has been determined at 2.6 Å resolution. The tetramer has approximate dihedral symmetry and each protomer consists of three structural domains with topologies similar to a jelly roll ...