Abstract:

Objective
To purify the phosphatidylethanolamine-binding protein 4 (PEBP4) , a new member of the phosphatidylethanolamine-binding protein family from  porcine seminal plasma and lay the foundation for further investigation of its function.Methods Sus PEBP4 was purified from approximately 500 mL porcine seminal plasma by ammonium sulfate fractionation, Q-Sepharose column chromatography, matrex Gel Red A column chromatography, SP-XL-Sepharose column chromatography and Mono-S(A) column chromatography (in FPLC).The amino acid sequence of the purified sus PEBP4 was identified by automated N-terminal sequencing of the intact protein.The amino acid sequences of sus PEBP4 were compared with those of human, bovine, dog PEBP4 by bioinformatics.Results The molecular mass of the purified protein was calculated to be 25 000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of beta-mercaptoethanol.The overall homology of amino acid sequence between sus, human, bovine and dog  was from 75.2 to 81.8%.Conclusion Sus PEBP4 is purified by the optimal methods from porcine seminal plasma.Its primary structure is defined.Sus PEBP4 has high homology with other organisms.

Key words: phosphatidylethanolamine-binding protein, purification, porcine seminal plasma, amino acid sequence