Biological activities of CNBr fragments of a major protein secreted from the rat seminal vesicle epithelium.
Two fragments of SV-IV, one of the major proteins secreted from the rat seminal vesicle epithelium, were produced in vitro by protein cleavage with CNBr at level of the single methionine residue (Met-70) occurring in its polypeptide chain. After their purification by reversed-phase chromatography, SV-IV/A (1-70 fragment) and SV-IV/B (71-90 fragment) were assayed as transglutaminase substrates, and their anti-inflammatory, anti-thrombotic and immunosuppressive properties were evaluated in comparison with native SV-IV. Both fragments retained the SV-IV ability to act as transglutaminase substrates in vitro; fast atom bombardment mass spectrometry analyses of the reaction products pointed to Gln-9 and Gln-86 as acyl donor sites, and to Lys-59, -79 and -80 as acyl acceptor sites. In contrast, only SV-IV/A was shown to possess, like SV-IV, the property of inhibiting both the intensity of the carrageenin-induced rat foot edema and the platelet aggregation induced in vivo by different agents. Finally, the two protein fragments were found to be completely unable to inhibit both the mitogen-induced proliferation of human T cells and the mixed lymphocyte reaction.
Porta
R
R
Department of Biochemistry and Biophysics, University of Naples, Italy.
Esposito
C
C
Schinina
M E
ME
Mancuso
F
F
Marino
G
G
Pucci
P
P
Peluso
G
G
Metafora
S
S
D000595:Amino Acid Sequence; D000818:Animals; D003488:Cyanogen Bromide / Q000378:metabolism*; D004848:Epithelium / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism* / Q000494:pharmacology*; D010449:Peptide Mapping; D029584:Prostatic Secretory Proteins; D011506:Proteins / Q000378:metabolism* / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D029589:Seminal Plasma Proteins; D012669:Seminal Vesicles / Q000378:metabolism*; D016339:Spectrometry, Mass, Fast Atom Bombardment; D011503:Transglutaminases / Q000378:metabolism
D010446:Peptide Fragments; D029584:Prostatic Secretory Proteins; D011506:Proteins; D029589:Seminal Plasma Proteins; C043076:beta-microseminoprotein; D011503:Transglutaminases; D003488:Cyanogen Bromide