The heterogeneity, mechanism of regulation and function of human mast cell tryptase
The heterogeneity, mechanism of regulation and function of human mast cell tryptase
The tetrameric protease tryptase, the most abundant protein product of human mast cells, is emerging as a key mediator of allergic disease and as a possible target for drug development. Although some heterogeneity has been reported, it has not been clear to what extent tryptase may represent a single enzyme or a family of related enzymes, or what association exists between conformation, enzymatic activity and function.
We have analysed the size and charge heterogeneity of tryptase, either purified from human tissues or present in lysates of purified lung and skin mast cells, by applying the sensitive technique of two-dimensional gel electrophoresis with Western blotting. Tryptase was predominantly in the form of dissociated monomers with 9 to 12 diffuse bands within the molecular weight range 29 to 40 kDa, and a pI or 5.1 to 6.3. However, in some preparations dimers, trimers and tetramers with or without degradation products were observed. Although the patterns obtained for lung and skin tryptase were broadly similar, there were small different between tissues and between individual donors. Lectin binding studies indicated the presence of complex-type oligosaccharide with varying degrees of sialylation. Deglycosylation with protein-N-glycosidase F reduced the size of both lung and skin tryptase, while incubation with PNGase F or neuraminidase resulted in a narrowing of the pI range, indicating that variable glycosylation contributes in a major way to the size and charge heterogeneity of tryptase.
This study has provided evidence that tryptase may be a key mediator of allergic disease and a promising target for therapeutic intervention. Cognisance will need to be taken of the extent of tryptase heterogeneity, but the development of selective inhibitors will not only aid understanding the roles of this major mast cell product, and could also provide a valuable new class of anti-allergic drugs.
University of Southampton
Peng, Qi
25d7ac63-3d4e-4700-bb39-3b2e3f6a9ab5
1998
Peng, Qi
25d7ac63-3d4e-4700-bb39-3b2e3f6a9ab5
Peng, Qi
(1998)
The heterogeneity, mechanism of regulation and function of human mast cell tryptase.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
The tetrameric protease tryptase, the most abundant protein product of human mast cells, is emerging as a key mediator of allergic disease and as a possible target for drug development. Although some heterogeneity has been reported, it has not been clear to what extent tryptase may represent a single enzyme or a family of related enzymes, or what association exists between conformation, enzymatic activity and function.
We have analysed the size and charge heterogeneity of tryptase, either purified from human tissues or present in lysates of purified lung and skin mast cells, by applying the sensitive technique of two-dimensional gel electrophoresis with Western blotting. Tryptase was predominantly in the form of dissociated monomers with 9 to 12 diffuse bands within the molecular weight range 29 to 40 kDa, and a pI or 5.1 to 6.3. However, in some preparations dimers, trimers and tetramers with or without degradation products were observed. Although the patterns obtained for lung and skin tryptase were broadly similar, there were small different between tissues and between individual donors. Lectin binding studies indicated the presence of complex-type oligosaccharide with varying degrees of sialylation. Deglycosylation with protein-N-glycosidase F reduced the size of both lung and skin tryptase, while incubation with PNGase F or neuraminidase resulted in a narrowing of the pI range, indicating that variable glycosylation contributes in a major way to the size and charge heterogeneity of tryptase.
This study has provided evidence that tryptase may be a key mediator of allergic disease and a promising target for therapeutic intervention. Cognisance will need to be taken of the extent of tryptase heterogeneity, but the development of selective inhibitors will not only aid understanding the roles of this major mast cell product, and could also provide a valuable new class of anti-allergic drugs.
This record has no associated files available for download.
More information
Published date: 1998
Identifiers
Local EPrints ID: 463615
URI: http://eprints.soton.ac.uk/id/eprint/463615
PURE UUID: e9db003b-1848-4ac5-ba13-f83999dc8880
Catalogue record
Date deposited: 04 Jul 2022 20:54
Last modified: 23 Jul 2022 02:15
Export record
Contributors
Author:
Qi Peng
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics