Communication between the nucleotide site and the main molecular hinge of 3-phosphoglycerate kinase.
3-Phosphoglycerate kinase is a hinge-bending enzyme with substrate-assisted domain closure. However, the closure mechanism has not been described in terms of structural details. Here we present experimental evidence of the participation of individual substrate binding side chains in the operation of the main hinge which is distant from the substrate binding sites. The combined mutational, kinetic, and structural (DSC and SAXS) data for human 3-phosphoglycerate kinase have shown that catalytic residue R38, which also binds the substrate 3-phosphoglycerate, is essential in inducing domain closure. Similarly, residues K219, N336, and E343 which interact with the nucleotide substrates are involved in the process of domain closure. The other catalytic residue, K215, covers a large distance during catalysis but has no direct role in domain closure. The transmission path of the nucleotide effect toward the main hinge of PGK is described for the first time at the level of interactions existing in the tertiary structure.
Institute of Enzymology, Biological Research Center, Hungarian Academy of Sciences, H-1518 Budapest, PO Box 7, Hungary, Germany.
Judit
J
Szabó
Andrea
A
Varga
Beáta
B
Flachner
Peter V
PV
Konarev
Dmitri I
DI
Svergun
Péter
P
Závodszky
Mária
M
Vas
D000818:Animals; D001665:Binding Sites; D002152:Calorimetry, Differential Scanning; D002384:Catalysis; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D006801:Humans; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D009711:Nucleotides / Q000737:chemistry* / Q000378:metabolism*; D010735:Phosphoglycerate Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D017186:Titrimetry; D044366:Transition Temperature; D014346:Trypanosoma brucei brucei / Q000201:enzymology
D009711:Nucleotides; D021122:Protein Subunits; D010735:Phosphoglycerate Kinase