Enzymatic action of prostate-specific antigen (PSA or hK3) : Substrate specificity and regulation by Zn2+, a tight-binding inhibitor
Malm, Johan LU ; Hellman, Jukka ; Hogg, Phil and Lilja, Hans LU
(2000)
In Prostate
45(2).
p.132-139
- Abstract
BACKGROUND. In semen, prostate-specific antigen (PSA or hK3) digests the gel proteins semenogelin I and II, resulting in liquefaction and the release of motile spermatozoa. We characterized the substrate specificity and zinc-mediated inhibition of PSA. METHODS. The proteolysis of human semenogelin I (SgI) and II (SgII) by PSA was characterized by purification of generated SgI and SgII fragments, N-terminal sequencing, and mass spectrometry. Zn2+-inhibition of PSA was studied using a chromogenic substrate. RESULTS. Eighteen cleavage sites in SgI and 16 in SgII were identified. Cleavages were identified mainly as the C-terminal of certain tyrosine and glutamine residues, but also the C-terminal of histidine, aspartic acid,... (More)
BACKGROUND. In semen, prostate-specific antigen (PSA or hK3) digests the gel proteins semenogelin I and II, resulting in liquefaction and the release of motile spermatozoa. We characterized the substrate specificity and zinc-mediated inhibition of PSA. METHODS. The proteolysis of human semenogelin I (SgI) and II (SgII) by PSA was characterized by purification of generated SgI and SgII fragments, N-terminal sequencing, and mass spectrometry. Zn2+-inhibition of PSA was studied using a chromogenic substrate. RESULTS. Eighteen cleavage sites in SgI and 16 in SgII were identified. Cleavages were identified mainly as the C-terminal of certain tyrosine and glutamine residues, but also the C-terminal of histidine, aspartic acid, leucine, serine, and asparagine residues. No cleavages were identified at any arginine, lysine, phenylalanine, tryptophan, or methionine residues, indicating that the substrate specificity of PSA is distinct from that of trypsin, chymotrypsin, tissue kallkrein (hK1), and kallikrein 2 (hK2). Zn2+ ions have a dramatic effect on PSA activity; the data indicate that Zn2+ is a tight-binding inhibitor of PSA activity. CONCLUSIONS. The data will enable the optimized design of PSA activity assays, which may prove instrumental to uncovering the role of PSA in cancer and reproduction. The inhibition data indicate that Zn2+ could regulate PSA activity, which may prove important in the development of efficient inhibitors of PSA activity. (C) 2000 Wiley-Liss, Inc.
(Less)
- author
- Malm, Johan
LU
; Hellman, Jukka
; Hogg, Phil
and Lilja, Hans
LU
- organization
- publishing date
- 2000
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Kallikrein, Prostate-specific antigen, Semenogelin, Substrate specificity, Zinc
- in
- Prostate
- volume
- 45
- issue
- 2
- pages
- 132 - 139
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:0033814795
- pmid:11027412
- ISSN
- 0270-4137
- DOI
- 10.1002/1097-0045(20001001)45:2<132::AID-PROS7>3.0.CO;2-3
- language
- English
- LU publication?
- yes
- id
- 3bc97c89-46cb-48c0-9b28-88c45a9e2637
- date added to LUP
- 2022-12-06 17:34:15
- date last changed
- 2024-09-15 18:48:04
@article{3bc97c89-46cb-48c0-9b28-88c45a9e2637,
abstract = {{<p>BACKGROUND. In semen, prostate-specific antigen (PSA or hK3) digests the gel proteins semenogelin I and II, resulting in liquefaction and the release of motile spermatozoa. We characterized the substrate specificity and zinc-mediated inhibition of PSA. METHODS. The proteolysis of human semenogelin I (SgI) and II (SgII) by PSA was characterized by purification of generated SgI and SgII fragments, N-terminal sequencing, and mass spectrometry. Zn<sup>2+</sup>-inhibition of PSA was studied using a chromogenic substrate. RESULTS. Eighteen cleavage sites in SgI and 16 in SgII were identified. Cleavages were identified mainly as the C-terminal of certain tyrosine and glutamine residues, but also the C-terminal of histidine, aspartic acid, leucine, serine, and asparagine residues. No cleavages were identified at any arginine, lysine, phenylalanine, tryptophan, or methionine residues, indicating that the substrate specificity of PSA is distinct from that of trypsin, chymotrypsin, tissue kallkrein (hK1), and kallikrein 2 (hK2). Zn<sup>2+</sup> ions have a dramatic effect on PSA activity; the data indicate that Zn<sup>2+</sup> is a tight-binding inhibitor of PSA activity. CONCLUSIONS. The data will enable the optimized design of PSA activity assays, which may prove instrumental to uncovering the role of PSA in cancer and reproduction. The inhibition data indicate that Zn<sup>2+</sup> could regulate PSA activity, which may prove important in the development of efficient inhibitors of PSA activity. (C) 2000 Wiley-Liss, Inc.</p>}},
author = {{Malm, Johan and Hellman, Jukka and Hogg, Phil and Lilja, Hans}},
issn = {{0270-4137}},
keywords = {{Kallikrein; Prostate-specific antigen; Semenogelin; Substrate specificity; Zinc}},
language = {{eng}},
number = {{2}},
pages = {{132--139}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Prostate}},
title = {{Enzymatic action of prostate-specific antigen (PSA or hK3) : Substrate specificity and regulation by Zn<sup>2+</sup>, a tight-binding inhibitor}},
url = {{http://dx.doi.org/10.1002/1097-0045(20001001)45:2<132::AID-PROS7>3.0.CO;2-3}},
doi = {{10.1002/1097-0045(20001001)45:2<132::AID-PROS7>3.0.CO;2-3}},
volume = {{45}},
year = {{2000}},
}