Bovine Seminal Protein Research Articles

Direct UV-MALDI-TOF MS Analysis of (Glyco)proteins of Fractions of Bovine Seminal Plasma

Bovine seminal plasma was submitted to chromatography on Con A-Sepharose. The “non-interacting”, “weakly-interacting” and “strongly-interacting” fractions were analyzed through UV-MALDI-TOF MS together with a subfraction of the “non-interacting” material, using sinapinic acid (SA) as matrix. The spectra were obtained in linear positive mode in the 4.0-90.0 kDa mass/charge range showing peaks in well defined zones, namely: 5.5-8.0 kDa, 10.0-12.0 kDa, 12.5-14.0 kDa (major), 23.2-23.7 kDa, 26.1-27.5 kDa and 38.0-40.0 kDa. High sensitivity spectra showed some very small peaks until 90 kDa. Bovine seminal protein (BSP-A3), acidic seminal fluid protein (αSFP) and PDC-109 glycoproteins (BSP-A1 and BSP-A2) were identified. Caltrin, the human epididymis-specific glycoprotein (HE4), the calcium transport inhibitor protein, the inhibitor of metalloprotease 2 (TIMP-2), osteopontin (OPN) and the prostatic acid protease (PAP) were tentatively identified. The molecular weight of some peaks can be arranged in a sequence from that of BSP-A3 going through the molecular weights of glycoforms (including the known BSP-A1 and BSP-A2) which differ in the amounts of neutral hexoses and sialic acids, composing a BSP-family more extended than previously reported. Another two families could be builded up from proteins of molecular weight of about 12730 and 12750 Da and glycoforms which differ from them also by hexoses and sialic acids. The structures of the deduced O-linked oligosaccharides of the glycoforms are in complete agreement to that determined for the BSP-A1 oligosaccharide. Small differences in the m. w. of some (glyco)proteins were attributed to genetic polymorphysm. The identification of proteins and O-linked glycoproteins in the “interacting” fractions of the chromatography suggests that the fractionation was not due to specific affinity interactions but to non-specific hydrophobic interactions of the proteins with the hydrophobic pocked of the Con A.

Effect of storage at 5 degrees C on the disc electrophoretic patterns of ovine and bovine seminal proteins.

Separation of ovine seminal plasma proteins in the disc electrophoretic system revealed a consistent pattern of sixteen fractions with some variation among six rams in the relative amount of minor components, whereas bovine seminal plasma had twelve fractions. In both species, the major seminal plasma components had electrophoretic mobilities somewhat less than did the albumin band of the corresponding blood plasma. The prealbumin region of the seminal plasma electrophoretograms contained four fractions in the ovine samples and three fractions in the bovine samples. Storage ofdiluted ovine and bovine semen at 5\s=deg\C did not alter the total protein concentration in the seminal plasma. Standard disc electrophoretic patterns showed that two of the sixteen ovine seminal plasma protein fractions disappeared during such storage. No changes were evident in either the standard or basic electrophoretograms of bovine seminal plasma during storage of semen. Standard disc electrophoretograms of ovine and bovine sperm extracts showed many lightly staining bands, but no progressive changes were evident in these electrophoretic patterns during storage of semen.