Adenosine deaminase from bovine brain: purification and partial characterization.
Bovine brain adenosine deaminase cytoplasmatic form was purified about 450 fold by salt fractionation, column chromatography on DEAE-cellulose, octyl-sepharose 4B and affinity chromatography on CH-sepharose 4B 9-(p-aminobenzyl)adenine. The purified enzyme was homogeneous on disc gel electrophoresis; the enzyme had a molecular mass of about 65 kDa with an isoelectric point at pH 4.87. The Km values for adenosine and 2'-deoxyadenosine were 4 x 10(-5) and 5.2 x 10(-5) M, respectively. The enzyme showed a great stability to temperature with a half life of 15 hours at 53 degrees C significantly different compared to that known for other mammalian forms of this enzyme. Aza and deaza analogs of adenosine and erythro-9-(2-hydroxy-3-nonyl) adenine were good inhibitors of the bovine brain enzyme with little difference with respect to those reported for the adenosine deaminases purified from other sources. Kinetic constants for the association and dissociation of coformycin and 2'-deoxycoformycin with the bovine brain adenosine deaminase are reported.
Lupidi
G
G
Dept. of M.C.A. Biology, University of Camerino, Italy.
Falasca
M
M
Marmocchi
F
F
Venardi
G
G
Cristalli
G
G
Riva
F
F
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000243:Adenosine Deaminase / Q000302:isolation & purification* / Q000378:metabolism; D058892:Adenosine Deaminase Inhibitors; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D003070:Coformycin / Q000378:metabolism / Q000494:pharmacology; D003593:Cytoplasm / Q000201:enzymology; D004795:Enzyme Stability; D066298:In Vitro Techniques; D007526:Isoelectric Point; D007700:Kinetics; D008970:Molecular Weight; D015649:Pentostatin / Q000378:metabolism / Q000494:pharmacology; D013696:Temperature
D058892:Adenosine Deaminase Inhibitors; D003070:Coformycin; D015649:Pentostatin; D000243:Adenosine Deaminase; D000241:Adenosine