Slow reversible inhibitions of rabbit muscle aldolase with substrate analogues: synthesis, enzymatic kinetics and UV difference spectroscopy studies.
Various dihydroxyacetone-phosphate (DHAP) analogues bearing an aromatic ring or beta-dicarbonyl structures were synthesized. Their capacity to form a stabilized iminium ion or conjugated enamine in the reaction catalyzed by rabbit muscle aldolase (EC 4.1.2.13) were investigated by enzymatic kinetics and UV difference spectroscopic techniques. Whereas the aromatic derivative led to competitive inhibition without detectable iminium ion formation, slow reversible inhibitions of aldolase by beta-dicarbonyl compounds was shown to have taken place. Conjugated enamine formation at the active site of the enzyme was detected by their specific absorbances close to 317 nm.
Gefflaut
T
T
Groupe de Chimie Organique Biologique, UMR 5623, Bát, IIR1, Université Paul Sabarier, Toulouse, France.
Blonski
C
C
Périé
J
J
D000098:Acetophenones / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D000614:Aminocaproates / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D004099:Dihydroxyacetone Phosphate / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology*; D005634:Fructose-Bisphosphate Aldolase / Q000037:antagonists & inhibitors*; D007700:Kinetics; D009132:Muscles / Q000201:enzymology; D010755:Organophosphates / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D010422:Pentanones / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D011817:Rabbits; D013057:Spectrum Analysis / Q000379:methods; D013997:Time Factors; D014466:Ultraviolet Rays
C105396:(2,4-dioxo)butyl phosphate; C105398:2-(2-hydroxyphenyl)-2-oxoethyl phosphate; D000098:Acetophenones; D000614:Aminocaproates; D004791:Enzyme Inhibitors; D010755:Organophosphates; D010422:Pentanones; C105397:1-(phosphonooxy)-2,4-pentanedione; D004099:Dihydroxyacetone Phosphate; D005634:Fructose-Bisphosphate Aldolase