Affinity chromatography of bull seminal proteins on mannan-Sepharose.
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.
Liberda
J
J
Department of Biochemistry, Charles University, Albertov 2030, 128 40 Praha 2, Czech Republic.
Ryslavá
H
H
Jelínková
P
P
Jonáková
V
V
Tichá
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D008351:Mannans / Q000737:chemistry; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012661:Semen / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry
D008351:Mannans; D011506:Proteins; D012685:Sepharose