Crystal structure of the W35A mutant thioredoxin h from Chlamydomonas reinhardtii: The substitution of the conserved active site Trp leads to modifications in the environment of the two catalytic cysteines
V. Menchise et al., Crystal structure of the W35A mutant thioredoxin h from Chlamydomonas reinhardtii: The substitution of the conserved active site Trp leads to modifications in the environment of the two catalytic cysteines, BIOPOLYMERS, 56(1), 2000, pp. 1-7
The conformational analysis of W35A thioredoxin h from the eukaryotic green
alga Chlamydomonas reinhardtii in the solid state has been carried out by
x-ray diffraction, with the aim to clarify the role of Trp in the catalysis
. Comparative analysis of W35A mutant with wild-type (WT) thioredoxin shows
that, even if the structural motif of thioredoxin is not perturbed, the su
bstitution of Trp35 by an Ala leads to significant changes in protein confo
rmation near the active site. This rearrangement increases its solvent expo
sure and explains the change of the pKa values of the catalytic cysteines.
The substitution of the Trp residue also influences the crystal packing as
well as the recognition ability of thioredoxin. The solid state analysis su
ggests that the Trp residue has a structural function both to force the act
ive site in the bioactive conformation, and to mediate the protein-protein
recognition. (C) 2001 John Wiley & Sons, Inc.