[Behavior of the enzymes of the salvage pathway of purine bases in leukemia lymphocytes].
The Authors present a procedure for the determination of adenine phosphoribosyltransferase (APRT) and hypoxanthine phosphoribosyltransferase (HPRT) in lymphocytes which exhibits high sensitivity and requires low quantities of lymphocytes. 5 normal subjects and 4 patients affected by chronic lymphocytic leukemia (CLL) were considered. Human lymphocytes were prepared and treated as previously reported. To the incubation mixtures buffered with 50 mM TRIS-HCl pH 7.4 either 14C-adenine or 14C-hypoxanthine was added: after deproteinization and neutralization we followed the formation of either 14C-adenylic acid (AMP) or 14C-inosinic acid (IMP) by HPLC. A Supelcosil C18 5 microns (250 X 4.5 mm) column was used: IMP was eluted with 20 mM KH2PO4 pH 5.5 while AMP with a linear gradient to 40% B in 20 min., where A was 20 mM KH2PO4 pH 5.5 and B methanol/water 60:40. Evaluation of AMP and IMP formed was carried out by determination of the radioactivity of the collected peaks. The values of APRT in leukemic patients were enhanced when referred to the proteins and those of HGPRT decreased: the Authors propose to complete the study evaluating the intracellular content of adenine and hypoxanthine.
Istituto di Chimica Biologica, Università di Siena.
A
A
Tabucchi
B
B
Porcelli
D
D
Vannoni
A
A
Di Stefano
M
M
Pizzichini
R
R
Leoncini
E
E
Dispensa
E
E
Marinello
D000228:Adenine Phosphoribosyltransferase / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000378:metabolism*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000201:enzymology*; D008214:Lymphocytes / Q000201:enzymology*; D010430:Pentosyltransferases / Q000378:metabolism*; D011687:Purines / Q000378:metabolism
D011687:Purines; D010430:Pentosyltransferases; D000228:Adenine Phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase
Comportamento degli enzimi della via di recupero delle basi puriniche in linfociti leucemici.