Reactivation of triosephosphate isomerase from three trypanosomatids and human: effect of suramin.
The reactivation of the homodimeric triosephosphate isomerases (TIMs) from Trypanosoma brucei, T. cruzi, Leishmania mexicana and humans was determined after their denaturation with guanidine hydrochloride. In the range of 2-32 microg of T. brucei TIM per ml and 0.2-5 microg of the other enzymes per ml, the rate and extent of TIM reactivation depended on protein concentration, indicating that at these protein concentrations, the rate-limiting step of reactivation is monomer association and not monomer folding. The rate of monomer association was more than one order of magnitude lower in the T. brucei enzyme than in the other three enzymes. Suramin is a drug of choice in the treatment of sleeping sickness, but its mechanism of action is not known. At micromolar concentrations, Suramin inhibited the reactivation of the four enzymes, but the extent of inhibition by Suramin decreased with increasing protein concentration as consequence of a diminution of the life time of the folded monomer. Since the life time of the monomer of T. brucei TIM is longer than that of the other enzymes, Suramin is a more effective inhibitor of the reactivation of TIM from T. brucei, particularly at monomer concentrations above 1 microg of protein per ml (monomer concentration approx. 37 nM). Compounds that are structurally related to Suramin also inhibit TIM reactivation; their effect was about five times more pronounced in the enzyme from T. brucei than in human TIM.
Gao
X G
XG
Departamento de Bioquímica, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Apartado Postal 70243, 04510 México, D.F., México.
Garza-Ramos
G
G
Saavedra-Lira
E
E
Cabrera
N
N
De Gómez-Puyou
M T
MT
Perez-Montfort
R
R
Gómez-Puyou
A
A
D000818:Animals; D004396:Coloring Agents / Q000494:pharmacology; D019281:Dimerization; D004789:Enzyme Activation / Q000187:drug effects; D019791:Guanidine / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D015394:Molecular Structure; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D013498:Suramin / Q000031:analogs & derivatives / Q000494:pharmacology*; D014305:Triose-Phosphate Isomerase / Q000378:metabolism*; D014345:Trypanosoma / Q000201:enzymology*
D004396:Coloring Agents; D013498:Suramin; D014305:Triose-Phosphate Isomerase; D019791:Guanidine