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Crystal structure of recombinant trypsin‐solubilized fragment of cytochrome b 5 and the structural comparison with Val61His mutant
Author(s) -
Wu Jian,
Gan JianHua,
Xia ZongXiang,
Wang YunHua,
Wang WenHu,
Xue LingLong,
Xie Yi,
Huang ZhongXian
Publication year - 2000
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/(sici)1097-0134(20000801)40:2<249::aid-prot70>3.0.co;2-h
Subject(s) - heme , chemistry , salt bridge , histidine , cytochrome , crystallography , protein structure , stereochemistry , mutant , biochemistry , amino acid , enzyme , gene
The crystal structure of the recombinant trypsin‐solubilized fragment of the microsomal cytochrome b 5 from bovine liver has been determined at 1.9 Å resolution and compared with the reported crystal structure of the lipase‐solubilized fragment of the membrane protein cytochrome b 5 . The two structures are similar to each other. However, some detailed structural differences are observed: the conformation of the segment Asn16–Ser20 is quite different, some helices around the heme and some segments between the helices are shifted slightly, the heme is rotated about the normal of the mean plane of heme, one of the propionates of the heme exhibits a different conformation. The average coordination distances between the iron and the two nitrogen atoms of the imidazole ligands are the same in the two structures. Most of the structural differences can be attributed to the different intermolecular interactions which result from the crystal packing. The wild‐type protein structure is also compared with its Val61His mutant, showing that the heme binding and the main chain conformations are basically identical with each other except for the local area of the mutation site. However, when Val61 is mutated to histidine, the large side chain of His61 is forced to point away from the heme pocket toward the solvent region, disturbing the micro‐environment of the heme pocket and influencing the stability and the redox potential of the protein. Proteins 2000;40:249–257. © 2000 Wiley‐Liss, Inc.