Donor substrate regulation of transketolase.
The influence of substrates on the interaction of apotransketolase with thiamin diphosphate was investigated in the presence of magnesium ions. It was shown that the donor substrates, but not the acceptor substrates, enhance the affinity of the coenzyme either to only one active center of transketolase or to both active centers, but to different degrees in each, resulting in a negative cooperativity for coenzyme binding. In the absence of donor substrate, negative cooperativity is not observed. The donor substrate did not affect the interaction of the apoenzyme with the inactive coenzyme analogue, N3'-pyridyl-thiamin diphosphate. The influence of the donor substrate on the coenzyme-apotransketolase interaction was predicted as a result of formation of the transketolase reaction intermediate 2-(alpha,beta-dihydroxyethyl)-thiamin diphosphate, which exhibited a higher affinity to the enzyme than thiamin diphosphate. The enhancement of thiamin diphosphate's affinity to apotransketolase in the presence of donor substrate is probably one of the mechanisms underlying the substrate-affected transketolase regulation at low coenzyme concentrations.
Esakova
Olga A
OA
A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia.
Meshalkina
Ludmilla E
LE
Golbik
Ralph
R
Hübner
Gerhard
G
Kochetov
German A
GA
D001665:Binding Sites; D002021:Buffers; D004305:Dose-Response Relationship, Drug; D015971:Gene Expression Regulation, Enzymologic; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007700:Kinetics; D008274:Magnesium / Q000737:chemistry; D015636:Magnesium Chloride / Q000737:chemistry; D008956:Models, Chemical; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013053:Spectrophotometry; D013696:Temperature; D013835:Thiamine Pyrophosphate / Q000737:chemistry; D013997:Time Factors; D014174:Transketolase / Q000737:chemistry* / Q000378:metabolism*
D002021:Buffers; D007477:Ions; D011994:Recombinant Proteins; D015636:Magnesium Chloride; D014174:Transketolase; D008274:Magnesium; D013835:Thiamine Pyrophosphate