A crystallographic study of the glutathione binding site of glutathione reductase at 0.3-nm resolution.
The binding of glutathione, some related molecules and two redox compounds to crystals of glutathione reductase has been investigated by X-ray crystallography at 0.3-nm resolution. Models for several bound ligands have been built and subjected to crystallographic refinement. The results clearly show the residues involved in glutathione binding as well as the geometry of the disulfide exchange. Glutathione-I is bound in a V-shaped conformation, while glutathione-II is extended. The zwitterionic glutamyl end of glutathione-II appears to be the most tightly bound part of the substrate. All glutathione conjugates and derivatives studied show binding dominated by the interactions at this site. In the reduced enzyme, glutathione-I forms a mixed disulfide intermediate with Cys58. Other structural changes are observed on reduction of the enzyme, and it is demonstrated that the carboxamidomethylated enzyme is a good model for the reduced species. Lipoate, a weak substrate of the enzyme, assumes a defined binding site where its disulfide is available for being attacked by Cys58-S gamma. A second region with affinity for a number of compounds has been found in a large cavity at the dimer interface of the enzyme. No functional role of this site is known.
Karplus
P A
PA
Institut für Organische Chemie und Biochemie der Universität Freiburg i. Br., Federal Rebpublic of Germany.
Pai
E F
EF
Schulz
G E
GE
D001665:Binding Sites; D004912:Erythrocytes / Q000201:enzymology; D005583:Fourier Analysis; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism*; D019803:Glutathione Disulfide; D005980:Glutathione Reductase / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D010619:Phenazines / Q000378:metabolism; D011487:Protein Conformation; D014812:Vitamin K / Q000378:metabolism; D014961:X-Ray Diffraction / Q000379:methods
D008024:Ligands; D010619:Phenazines; D014812:Vitamin K; D005980:Glutathione Reductase; D005978:Glutathione; D019803:Glutathione Disulfide; C009195:safranine T