A simple and efficient method for the purification of seminal plasmin, an antimicrobial protein from bovine seminal plasma.
A method involving direct affinity chromatography of undialysed bovine seminal plasma on a calmodulin-agarose column was developed for the purification of seminalplasmin. Seminalplasmin thus obtained was further purified from any contaminants left by ion-exchange chromatography on a short CM-Sepharose column. The method gave an excellent yield of seminalplasmin (0.7 mg per ml seminal plasma) that showed 100% inhibition of bacterial growth at a concentration of 10 micrograms/ml. The purified seminalplasmin was found to be homogeneous as tested by HPLC on a reverse-phase column and SDS-PAGE.
Kameswari
D B
DB
Centre for Cellular & Molecular Biology, Hyderabad.
Prasad
K S
KS
D000818:Animals; D000890:Anti-Infective Agents / Q000302:isolation & purification*; D002147:Calmodulin; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D011506:Proteins / Q000302:isolation & purification*; D012661:Semen / Q000737:chemistry*; D029607:Seminal Vesicle Secretory Proteins; D012685:Sepharose
D000890:Anti-Infective Agents; D002147:Calmodulin; D011506:Proteins; D029607:Seminal Vesicle Secretory Proteins; D012685:Sepharose