Kinetic investigation of cooperativity in coenzyme binding by transketolase active sites.
The two-step mechanism of coenzyme (thiamine diphosphate, ThDP) binding with two initially identical active sites of apotransketolase has been examined with a kinetic model. Cooperativity between sites in the primary ThDP binding and in the following conformational transition has been analyzed. The only reliable difference between sites is shown to be the tenfold difference in the backward rate constants of the conformational transition; this means that the cooperative interaction between sites takes place only after termination of both steps of ThDP binding in both sites.
Kovina
M V
MV
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119899, Russia.
Selivanov
V A
VA
Kochevova
N V
NV
Kochetov
G A
GA
D001051:Apoenzymes / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D007700:Kinetics; D008956:Models, Chemical; D008962:Models, Theoretical; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013835:Thiamine Pyrophosphate / Q000378:metabolism*; D014174:Transketolase / Q000737:chemistry* / Q000378:metabolism*
D001051:Apoenzymes; D014174:Transketolase; D013835:Thiamine Pyrophosphate