Functional proteomic profiling reveals KLK13 and TMPRSS11D as active proteases in the lower female reproductive tract [version 2; referees: 2 approved]

Background: Cervical-vaginal fluid (CVF) hydrates the mucosa of the lower female reproductive tract and is known to contain numerous proteases. The low pH of CVF (4.5 or below in healthy women of reproductive age) is a uniquely human attribute and poses a challenge for the proteolytic functioning of...

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Main Authors: Carla M.J. Muytjens, Yijing Yu, Eleftherios P. Diamandis
Format: Article
Language:English
Published: F1000 Research Ltd 2018-12-01
Series:F1000Research
Online Access:https://f1000research.com/articles/7-1666/v2
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author Carla M.J. Muytjens
Yijing Yu
Eleftherios P. Diamandis
author_facet Carla M.J. Muytjens
Yijing Yu
Eleftherios P. Diamandis
author_sort Carla M.J. Muytjens
collection DOAJ
description Background: Cervical-vaginal fluid (CVF) hydrates the mucosa of the lower female reproductive tract and is known to contain numerous proteases. The low pH of CVF (4.5 or below in healthy women of reproductive age) is a uniquely human attribute and poses a challenge for the proteolytic functioning of the proteases identified in this complex biological fluid. Despite the abundance of certain proteases in CVF, the proteolytic activity and function of proteases in CVF is not well characterized. Methods: In the present study, we employed fluorogenic substrate screening to investigate the influence of pH and inhibitory compounds on the proteolytic activity in CVF. Activity-based probe (ABP) proteomics has evolved as a powerful tool to investigate active proteases within complex proteomes and a trypsin-specific ABP was used to identify active proteases in CVF. Results: Serine proteases are among the most abundant proteins in the CVF proteome. Labeling human CVF samples with the trypsin-specific ABP revealed serine proteases transmembrane protein serine 11D and kallikrein-related peptidase 13 as active proteases in CVF. Furthermore, we demonstrated that the proteolytic activity in CVF is highly pH-dependent with an almost absolute inhibition of trypsin-like proteolytic activity at physiological pH levels. Conclusions: These findings provide a framework to understand proteolytic activity in CVF. Furthermore, the present results provide clues for a novel regulatory mechanism in which fluctuations in CVF pH have the potential to control the catalytic activity in the lower female reproductive tract.
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spelling doaj.art-db74f911e1a041f7a316ff76186bf0712022-12-21T18:30:35ZengF1000 Research LtdF1000Research2046-14022018-12-01710.12688/f1000research.16255.219125Functional proteomic profiling reveals KLK13 and TMPRSS11D as active proteases in the lower female reproductive tract [version 2; referees: 2 approved]Carla M.J. Muytjens0Yijing Yu1Eleftherios P. Diamandis2Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, CanadaDepartment of Clinical Biochemistry, University Health Network, Canada, Toronto, CanadaBackground: Cervical-vaginal fluid (CVF) hydrates the mucosa of the lower female reproductive tract and is known to contain numerous proteases. The low pH of CVF (4.5 or below in healthy women of reproductive age) is a uniquely human attribute and poses a challenge for the proteolytic functioning of the proteases identified in this complex biological fluid. Despite the abundance of certain proteases in CVF, the proteolytic activity and function of proteases in CVF is not well characterized. Methods: In the present study, we employed fluorogenic substrate screening to investigate the influence of pH and inhibitory compounds on the proteolytic activity in CVF. Activity-based probe (ABP) proteomics has evolved as a powerful tool to investigate active proteases within complex proteomes and a trypsin-specific ABP was used to identify active proteases in CVF. Results: Serine proteases are among the most abundant proteins in the CVF proteome. Labeling human CVF samples with the trypsin-specific ABP revealed serine proteases transmembrane protein serine 11D and kallikrein-related peptidase 13 as active proteases in CVF. Furthermore, we demonstrated that the proteolytic activity in CVF is highly pH-dependent with an almost absolute inhibition of trypsin-like proteolytic activity at physiological pH levels. Conclusions: These findings provide a framework to understand proteolytic activity in CVF. Furthermore, the present results provide clues for a novel regulatory mechanism in which fluctuations in CVF pH have the potential to control the catalytic activity in the lower female reproductive tract.https://f1000research.com/articles/7-1666/v2
spellingShingle Carla M.J. Muytjens
Yijing Yu
Eleftherios P. Diamandis
Functional proteomic profiling reveals KLK13 and TMPRSS11D as active proteases in the lower female reproductive tract [version 2; referees: 2 approved]
F1000Research
title Functional proteomic profiling reveals KLK13 and TMPRSS11D as active proteases in the lower female reproductive tract [version 2; referees: 2 approved]
title_full Functional proteomic profiling reveals KLK13 and TMPRSS11D as active proteases in the lower female reproductive tract [version 2; referees: 2 approved]
title_fullStr Functional proteomic profiling reveals KLK13 and TMPRSS11D as active proteases in the lower female reproductive tract [version 2; referees: 2 approved]
title_full_unstemmed Functional proteomic profiling reveals KLK13 and TMPRSS11D as active proteases in the lower female reproductive tract [version 2; referees: 2 approved]
title_short Functional proteomic profiling reveals KLK13 and TMPRSS11D as active proteases in the lower female reproductive tract [version 2; referees: 2 approved]
title_sort functional proteomic profiling reveals klk13 and tmprss11d as active proteases in the lower female reproductive tract version 2 referees 2 approved
url https://f1000research.com/articles/7-1666/v2
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