Aj. Ketterman et al., Single amino acid changes outside the active site significantly affect activity of glutathione S-transferases, INSEC BIO M, 31(1), 2001, pp. 65-74
Glutathione S-transferases (GSTs: E.C. 2.5.1.18) are a multigene family of
multifunctional dimeric proteins that play a central role in detoxication.
Four allelic forms of the mosquito Anopheles dirus GST, adGST1-1, were clon
ed, expressed and characterized. The one or two amino acid changes in each
allelic form was shown to confer different kinetic properties. Based on an
available crystal structure, several of the residue changes were not in the
putative substrate-binding pocket. Modeling showed that these insect Delta
class GSTs also possess a hydrophobic surface pocket reported for Alpha, M
u and Pi class GSTs. The atom movement after replacement and minimization s
howed an average atom movement of about 0.1 Angstrom for the 0 to 25 Angstr
om distance from the alpha carbon of the single replaced residue. This does
not appear to be a significant movement in a static modeled protein struct
ure. However, 200-500 atoms were involved with movements greater than 0.2 A
ngstrom. Dynamics simulations were performed to study the effects this phen
omenon would exert on the accessible conformations. The data show that resi
dues affecting nearby responsive regions of tertiary structure can modulate
enzyme specificities, possibly through regulating attainable configuration
s of the protein. (C) 2001 Elsevier Science Ltd. All rights reserved.