Expression of bovine seminal ribonuclease in Escherichia coli.
Bovine seminal RNAase (BS-RNAase), an unusually dimeric member of the pancreatic-like ribonuclease superfamily, is also a multifunctional biological effector, with antitumor, immunosuppressive, and antispermatogenic activities. We report here the cloning of a semi-synthetic cDNA coding for the protein subunit chain, its expression with a T7 expression system in Escherichia coli inclusion bodies, the dimerization of correctly reoxidized monomeric protein, followed by the purification in high yields of the recombinant enzyme, and by its conversion to a protein undistinguishable from BS-RNAase as isolated from seminal vesicles, both in its catalytic activity and in the micro-heterogeneity of its quaternary structure.
Dipartimento di Chimica Organica e Biologica, Università di Napoli Federico II, Italy.
M
M
de Nigris
N
N
Russo
R
R
Piccoli
G
G
D'Alessio
A
A
Di Donato
D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012260:Ribonucleases / Q000235:genetics* / Q000302:isolation & purification; D012661:Semen / Q000201:enzymology*
D011994:Recombinant Proteins; D004247:DNA; D012260:Ribonucleases