Bull seminal plasma phosphodiesterase. Purification and general properties.
A phosphodiesterase from bull seminal plasma was purified to homogeneity. The purification procedure involved sequential column chromatographies on DEAE-Sephadex A-50, ConA-Agarose, chromatofocusing and AMP-Agarose. The final yield was about 20% with a 3000-fold purification. As indicated by chromatofocusing, the enzyme is an acidic protein (pI approximately 4.6) and owing to its interaction with Concanavalin A it is also a glycoprotein. The SDS-PAGE showed that the purified phosphodiesterase seemed to be constituted of a single polypeptide chain of about 125 kDa. The enzyme did not show an absolute substrate specificity. Thus, it was able to hydrolyze 4-nitrophenyl ester of 5'-TMP (but not of 3'-TMP), cAMP, nucleic acids as well as NAD+, ADP and ATP. According to its enzymatic properties, bull seminal plasma phosphodiesterase is to be considered an oligonucleate 5'-nucleotidohydrolase. In addition the seminal plasma phosphodiesterase also showed phosphonate esterase activity.
Codini
M
M
Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università di Perugia, Italy.
Fini
C
C
Paolotti
P
P
Floridi
A
A
D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008297:Male; D010727:Phosphoric Diester Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D012661:Semen / Q000201:enzymology*; D013379:Substrate Specificity
D010727:Phosphoric Diester Hydrolases