Putrescine activated S-adenosylmethionine decarboxylase from Trypanosoma brucei brucei.
Trypanosoma brucei brucei contained a S-adenosyl-L-methionine decarboxylase (AdoMetDC) strongly activated by putrescine. The enzyme was also activated to a lesser extent by cadaverine and 1,3-diaminopropane. Spermidine and spermine had no effect on basal activity of the enzyme. However, they interfered with putrescine activation of trypanosomal AdoMetDC. The trypanosomal enzyme could not be precipitated with antiserum against human AdoMetDC. The trypanosomal AdoMetDC enzyme subunit was labeled by reaction with 35S-decarboxylated AdoMet in the presence of NaCNBH4, and found to have a molecular weight of 34 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The subunit was readily degraded on storage to a form with a molecular weight of 26 kDa. The specificity of labeling of AdoMetDC by this procedure was confirmed by the prevention of 35S-decarboxylated S-adenosylmethionine (AdoMet) binding in the presence of specific AdoMetDC inhibitors [either methylglyoxal bis(guanylhydrazone (MGBG), a reversible inhibitor, or 5'-deoxy-5'-[(2-hydrazinoethyl)methylamino]adenosine (MHZEA), an irreversible inactivator]. As compared to human AdoMetDC, the trypanosomal enzyme showed weaker binding to a column of MGBG-Sepharose and also was significantly less sensitive to inhibition by MGBG and its congener ethylglyoxal bis(guanylhydrazone) (EGBG). Thus, the trypanosomal AdoMetDC differs significantly from its mammalian and bacterial counterparts and may therefore be exploited as a specific target for chemotherapy of trypanosomiasis.
Tekwani
B L
BL
Department of Cellular & Molecular Physiology, Milton S. Hershey Medical Center Pennsylvania State University College of Medicine, Hershey 17033.
Bacchi
C J
CJ
Pegg
A E
AE
D012437:Adenosylmethionine Decarboxylase / Q000037:antagonists & inhibitors / Q000187:drug effects* / Q000302:isolation & purification; D000818:Animals; D015317:Biogenic Polyamines / Q000494:pharmacology; D004789:Enzyme Activation; D005260:Female; D008322:Mammals / Q000378:metabolism / Q000469:parasitology; D008935:Mitoguazone / Q000031:analogs & derivatives / Q000494:pharmacology; D008970:Molecular Weight; D011233:Precipitin Tests; D011700:Putrescine / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D013462:Sulfur Radioisotopes; D014346:Trypanosoma brucei brucei / Q000201:enzymology*
D015317:Biogenic Polyamines; D013462:Sulfur Radioisotopes; C041127:ethylglyoxal bis(guanylhydrazone); D012437:Adenosylmethionine Decarboxylase; D008935:Mitoguazone; D011700:Putrescine