Isolation and characterization of the major gel proteins in human semen, semenogelin I and semenogelin II.
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Malm J, Hellman J, Magnusson H, Laurell CB, Lilja H
Isolation and characterization of the major gel proteins in human semen, semenogelin I and semenogelin II.
Eur J Biochem. 1996 May 15;238(1):48-53. doi: 10.1111/j.1432-1033.1996.0048q.x.
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- 8665951 [ View in PubMed]
- Abstract
Semenogelin I and semenogelin II constitute the major gel-forming proteins in human semen. The gel proteins were rapidly solubilized and separated from spermatozoa in ejaculates collected at pH 9.7 in buffer containing 4 mol/l urea and dithiothreitol. This protected the semenogelins from proteolytic degradation by prostate-specific antigen, and allowed their isolation by affinity chromatography on heparin-Sepharose. Semenogelins I and II were almost selectively retained and eluted partially separated in 0.25 mol/l NaCl. Further purification was achieved by chromatography on Superose. Approximately 10-20 mg semenogelin I and 2-5 mg semenogelin II were recovered from each sample with a purity exceeding 95% as judged by SDS/PAGE. The molecular mass of semenogelin I (49 958 Da) and the major form of semenogelin II (63 539 Da) measured by mass spectrometry was consistent with the reported cDNA data. The occurrence of a second, larger form of semenogelin II was due to asparagine-linked glycosylation. The amino-termini of the purified proteins were blocked, but digestion with pyroglutamate amino-peptidase enabled the identification of amino-terminal sequences consistent with the reported cDNA data. The amino acid compositions of the purified proteins were also consistent with those derived from cDNA data. The absorption coefficients (280 nm, 1%, 1 cm) for semenogelins I and II were 5.5 and 5.4, respectively, and the isoelectric point was above pH 9.5 for both proteins.