Isolation and characterization of the major gel proteins in human semen, semenogelin I and semenogelin II
Malm, Johan LU ; Hellman, Jukka ; Magnusson, Helena ; Laurell, Carl Bertil LU and Lilja, Hans LU
(1996)
In European Journal of Biochemistry
238(1).
p.48-53
- 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 chromatograph:g 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... (More)
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 chromatograph:g 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 aminopeptidase 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 semenoaelins I and II. were 5.5 and 5.4, respectively, and the isoelectric point was above pH 9.5 for both proteins.
(Less)
- author
- Malm, Johan
LU
; Hellman, Jukka
; Magnusson, Helena
; Laurell, Carl Bertil
LU
and Lilja, Hans
LU
- organization
- publishing date
- 1996
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Semenogelin, Seminal coagulum, Seminal plasma, Seminal vesicle
- in
- European Journal of Biochemistry
- volume
- 238
- issue
- 1
- pages
- 48 - 53
- publisher
- Wiley-Blackwell
- external identifiers
-
- scopus:0029882990
- pmid:8665951
- ISSN
- 0014-2956
- DOI
- 10.1111/j.1432-1033.1996.0048q.x
- language
- English
- LU publication?
- yes
- id
- d98dd28a-1da8-46f6-81fe-672cba8b692c
- date added to LUP
- 2022-12-06 16:37:59
- date last changed
- 2024-04-04 12:14:09
@article{d98dd28a-1da8-46f6-81fe-672cba8b692c,
abstract = {{<p>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 chromatograph:g 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 aminopeptidase 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 semenoaelins I and II. were 5.5 and 5.4, respectively, and the isoelectric point was above pH 9.5 for both proteins.</p>}},
author = {{Malm, Johan and Hellman, Jukka and Magnusson, Helena and Laurell, Carl Bertil and Lilja, Hans}},
issn = {{0014-2956}},
keywords = {{Semenogelin; Seminal coagulum; Seminal plasma; Seminal vesicle}},
language = {{eng}},
number = {{1}},
pages = {{48--53}},
publisher = {{Wiley-Blackwell}},
series = {{European Journal of Biochemistry}},
title = {{Isolation and characterization of the major gel proteins in human semen, semenogelin I and semenogelin II}},
url = {{http://dx.doi.org/10.1111/j.1432-1033.1996.0048q.x}},
doi = {{10.1111/j.1432-1033.1996.0048q.x}},
volume = {{238}},
year = {{1996}},
}