Affinity chromatography of bull seminal proteins on mannan–Sepharose

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2002-11, Vol.780 (2), p.231-239
• Main Authors: Liberda, J, Ryšlavá, H, Jelı́nková, P, Jonáková, V, Tichá, M
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Cattle; Chromatography, Affinity - methods; Electrophoresis, Polyacrylamide Gel; Male; Mannan; Mannans - chemistry; Proteins; Proteins - chemistry; Proteins - isolation & purification; Semen - chemistry; Sepharose - chemistry
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/S1570-0232(02)00521-4

The interaction of bull seminal plasma proteins and sperm with mannan was investigated using an enzyme-linked binding assay (ELBA). A high mannan-binding activity was found in the protein fraction interacting with heparin. Mannan binding to seminal plasma proteins was inhibited by d-mannose and d-fructose, but not by d-mannose-6-phosphate, d-glucose-6-phosphate, ovalbumin and ovomucoid. Mannan inhibited the binding of bovine zona pellucida glycoproteins both to bull sperm and seminal plasma proteins. Yeast mannan immobilized to divinyl sulfone-activated Sepharose was used for the isolation of mannan-binding proteins. The protein components of this fraction were identified on the basis of relative molecular mass determination and N-terminal amino acid sequencing: RNAase dimer, PDC-109 and a protein homologous to BSP-30K (relative molecular mass 14 500). The isolated proteins were characterized by a high zona pellucida binding activity.