Substrate specificity of the purine-2'-deoxyribonucleosidase of Crithidia luciliae.
The purine-2'-deoxyribonucleosidase of Crithidia luciliae catalyses an efficient deoxyribosyl transfer between a variety of purine bases, benzimidazole and 5,6-dimethylbenzimidazole. Since the deoxyriboside of a deoxyribosyl acceptor is necessarily also a substrate, the trans-N-deoxyribosylase activity of the enzyme allows a study of its specificity to be extended to a large number of purines and purine analogues. Amongst 27 different deoxyribosyl acceptors, only hypoxanthine gave rise to isomeric products. The introduction of methyl groups at appropriate positions in either purine or benzimidazole lowered the Michaelis constant, KB, for deoxyribosyl acceptors: by about 10-fold for 6-methylpurine (KB 351 +/- 87 microM) compared with purine (KB 3.91 +/- 0.8 mM) and by about 10(3)-fold for 5,6-dimethylbenzimidazole (KB 7.0 +/- 0.79 microM) compared with benzimidazole (Km,app. 7.8 +/- 2.4 mM). The maximal rates of deoxyribosyl transfer to different acceptors, on the other hand, varied by only 4.5-fold, and can be ascribed to decreases in the rate of release of the newly formed purine deoxyriboside from the enzyme.
Steenkamp
D J
DJ
Department of Chemical Pathology, University of Cape Town Medical School, South Africa.
Hälbich
T J
TJ
D000818:Animals; D003421:Crithidia / Q000201:enzymology*; D003839:Deoxyadenosines / Q000378:metabolism; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000378:metabolism; D007288:Inosine / Q000031:analogs & derivatives / Q000378:metabolism; D010430:Pentosyltransferases / Q000378:metabolism*; D013379:Substrate Specificity
D003839:Deoxyadenosines; D007042:Hypoxanthines; D019271:Hypoxanthine; D007288:Inosine; D010430:Pentosyltransferases; C021888:nucleoside deoxyribosyltransferase; C012271:deoxyinosine