Correlation between enzyme activity and hinge-bending domain displacement in 3-phosphoglycerate kinase.
Diffuse X-ray-scattering data give evidence for large-scale structural change in pig muscle 3-phosphoglycerate kinase upon substrate binding. Simultaneous binding of 3-phosphoglycerate and MgATP either to the unmodified enzyme or to its active methylated derivative leads to about an 0.1-nm decrease in radius of gyration. These data coincide well with the previous data for yeast 3-phosphoglycerate kinase. When, instead of methylation, the two reactive thiol groups of pig muscle 3-phosphoglycerate kinase are carboxamidomethylated, the enzyme becomes inactive and the radii of gyration of its 'apo' and 'holo' forms do not differ within limits of experimental error. Thus, a correlation exists between the activity of 3-phosphoglycerate kinase and its substrate-induced large-scale conformational change. This correlation is a strong argument in favor of the functional importance of domain locking in the reaction catalyzed by 3-phosphoglycerate kinase.
Sinev
M A
MA
Institute of Protein Research, Academy of Sciences of the USSR, Pushchino, Moscow Region.
Razgulyaev
O I
OI
Vas
M
M
Timchenko
A A
AA
Ptitsyn
O B
OB
D000478:Alkylation; D000818:Animals; D001665:Binding Sites; D008745:Methylation; D009132:Muscles / Q000201:enzymology*; D010735:Phosphoglycerate Kinase / Q000378:metabolism*; D011487:Protein Conformation; D012542:Scattering, Radiation; D013379:Substrate Specificity; D013552:Swine; D014961:X-Ray Diffraction; D015003:Yeasts / Q000201:enzymology
D010735:Phosphoglycerate Kinase