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Studies on a serine peptidase inhibitor locus on human chromosome 20 Characterization of novel genes encoding proteins with WFDC and kunitz domains

Clauss, Adam LU (2006) In Faculty of Medicine Doctoral Dissertation Series 2006:86.
Abstract
An unlimited number of proteolytic reactions are carried out by enzymes in biological systems. These enzymes are a necessity of life, but they can also be dangerous if they are not controlled. Protease inhibitors regulate proteolytic enzymes by interacting and forming complexes with them, thereby preventing the substrate from gaining access to the active site of the enzyme. We have recently identified a locus on human chromosome 20 that encompasses several genes that encode proteins containing one or more of the WFDC and kunitz serine protease domains. Among these genes are the previously described PI3 and SLPI. These new genes were first detected by their sequence similarity to the genes that encode the predominant gel-forming proteins in... (More)
An unlimited number of proteolytic reactions are carried out by enzymes in biological systems. These enzymes are a necessity of life, but they can also be dangerous if they are not controlled. Protease inhibitors regulate proteolytic enzymes by interacting and forming complexes with them, thereby preventing the substrate from gaining access to the active site of the enzyme. We have recently identified a locus on human chromosome 20 that encompasses several genes that encode proteins containing one or more of the WFDC and kunitz serine protease domains. Among these genes are the previously described PI3 and SLPI. These new genes were first detected by their sequence similarity to the genes that encode the predominant gel-forming proteins in semen, semenogelin I and semenogelin II. To evaluate the function of these novel proteins, we used RT-PCR to study their expression pattern in a panel of tissues and cell lines. Most genes are ubiquitously expressed, although higher levels of expression have been detected in the male reproductive organs, including the seminal vesicles, epididymis, testes, and prostate. Expression has also been observed in the LNCaP and PC3 prostate cancer cell lines, the former of which is androgen sensitive and the latter androgen independent. The semenogelin genes have previously been described to be rapidly evolving. We identified and compared the locus of four mammalian species, and the results demonstrate that the genes are under evolutionary pressure, which is consistent with those involved in reproduction and/or immune defence. Three recombinant proteins were constructed in a prokaryote system to study their anti-proteolytic capacity. We did not detect any inhibitory effect on a panel of enzymes, possibly due to misfolding of the recombinant proteins. To circumvent this problem, we plan to construct new recombinant proteins in a eukaryotic system to ensure correct folding and disulphide paring. (Less)
Please use this url to cite or link to this publication:
author
supervisor
opponent
  • Professor Ny, Tor, Umeå University
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Medicin (människa och djur), Medicine (human and vertebrates), kunitz, Protase inhibitor, WFDC
in
Faculty of Medicine Doctoral Dissertation Series
volume
2006:86
pages
93 pages
publisher
Adam Clauss Department of Laboratory Medicine Division of Clinical Chemistry University Hospital MAS Lund University S-205 02 Malmö, Sweden
defense location
Jubileumsaulan, Medicinskt forskningscentrum, Ing. 59, Universitetssjukhuset MAS
defense date
2006-05-24 09:15:00
ISSN
1652-8220
ISBN
91-85559-10-5
language
English
LU publication?
yes
additional info
id
9deccaa9-66ad-44c7-8339-f900ae492114 (old id 546741)
date added to LUP
2016-04-01 16:49:54
date last changed
2019-05-21 14:22:05
@phdthesis{9deccaa9-66ad-44c7-8339-f900ae492114,
  abstract     = {An unlimited number of proteolytic reactions are carried out by enzymes in biological systems. These enzymes are a necessity of life, but they can also be dangerous if they are not controlled. Protease inhibitors regulate proteolytic enzymes by interacting and forming complexes with them, thereby preventing the substrate from gaining access to the active site of the enzyme. We have recently identified a locus on human chromosome 20 that encompasses several genes that encode proteins containing one or more of the WFDC and kunitz serine protease domains. Among these genes are the previously described PI3 and SLPI. These new genes were first detected by their sequence similarity to the genes that encode the predominant gel-forming proteins in semen, semenogelin I and semenogelin II. To evaluate the function of these novel proteins, we used RT-PCR to study their expression pattern in a panel of tissues and cell lines. Most genes are ubiquitously expressed, although higher levels of expression have been detected in the male reproductive organs, including the seminal vesicles, epididymis, testes, and prostate. Expression has also been observed in the LNCaP and PC3 prostate cancer cell lines, the former of which is androgen sensitive and the latter androgen independent. The semenogelin genes have previously been described to be rapidly evolving. We identified and compared the locus of four mammalian species, and the results demonstrate that the genes are under evolutionary pressure, which is consistent with those involved in reproduction and/or immune defence. Three recombinant proteins were constructed in a prokaryote system to study their anti-proteolytic capacity. We did not detect any inhibitory effect on a panel of enzymes, possibly due to misfolding of the recombinant proteins. To circumvent this problem, we plan to construct new recombinant proteins in a eukaryotic system to ensure correct folding and disulphide paring.},
  author       = {Clauss, Adam},
  isbn         = {91-85559-10-5},
  issn         = {1652-8220},
  language     = {eng},
  publisher    = {Adam Clauss Department of Laboratory Medicine Division of Clinical Chemistry University Hospital MAS Lund University S-205 02 Malmö, Sweden},
  school       = {Lund University},
  series       = {Faculty of Medicine Doctoral Dissertation Series},
  title        = {Studies on a serine peptidase inhibitor locus on human chromosome 20 Characterization of novel genes encoding proteins with WFDC and kunitz domains},
  volume       = {2006:86},
  year         = {2006},
}