Purification and characterization of fertility-associated antigen (FAA) in bovine seminal fluid.
Heparin-binding proteins (HBP) recognized by a monoclonal antibody (M1) are produced by male accessory sex glands and bind to distinct regions of ejaculated bull sperm. Immunoblots of sperm proteins probed with M1 identified HBP variants of approximately 31-, 24-, and 21.5-kDa that were associated with increased fertility of bulls. The purpose of this study was to identify the 31-kDa HBP known as fertility-associated antigen (FAA). FAA was isolated by heparin-affinity chromatography and reversed-phase high performance liquid chromatography near homogeneity. Biochemical characterization indicated that FAA was an unglycosylated, basic protein. FAA protein was detected in seminal vesicle and prostate gland homogenates, and FAA extracted from sperm membranes by treatment with hypertonic media was identical biochemically to seminal fluid-derived FAA. N-terminal sequence analysis of purified FAA yielded a 26 amino acid sequence (L K I X S F N V R S F G E S K K A G F N A M R V I V) with 73% identity to a recently identified human deoxyribonuclease (DNase) I-like protein. Two internal amino acid sequences generated from lys-C digested FAA were 85% and 92% identical to the same DNase I-like protein. In conclusion, we have identified a bovine seminal heparin-binding protein that binds to sperm and is indicative of bull fertility as being similar to the family of DNase I-like proteins. These data demonstrate the presence of a novel DNase I-like protein in bull accessory sex glands and form the groundwork for the identification of a candidate genetic marker for fertility of bulls.
McCauley
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TC
Department of Animal Sciences, University of Arizona, Tucson, Arizona 85721-0038, USA.
Zhang
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H
Bellin
M E
ME
Ax
R L
RL
D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005299:Fertility Agents; D005303:Fertility Agents, Male / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D015151:Immunoblotting; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D008566:Membranes / Q000378:metabolism; D008969:Molecular Sequence Data; D011467:Prostate / Q000276:immunology / Q000378:metabolism; D012661:Semen / Q000737:chemistry*; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013094:Spermatozoa / Q000276:immunology / Q000378:metabolism
D000954:Antigens, Surface; D005299:Fertility Agents; D005303:Fertility Agents, Male; C408753:Fertility-associated antigen, Bos taurus; D008562:Membrane Glycoproteins