MMPs (matrix metalloproteases) are a family of zinc-dependent endopeptidases widely distributed throughout all kingdoms of life. In mammals, MMPs play key roles in many physiological and pathological processes, including remodelling of the extracellular matrix. In the genome of the annual plant Arabidopsis thaliana, five MMP-like proteins (At-MMPs) are encoded, but their function is unknown. Previous work on these enzymes was limited to gene expression analysis, and so far proteolytic activity has been shown only for At1-MMP. We expressed and purified the catalytic domains of all five At-MMPs as His-tagged proteins in Escherichia coli cells to delineate the biochemical differences and similarities among the Arabidopsis MMP family members. We demonstrate that all five recombinant At-MMPs are active proteases with distinct preferences for different protease substrates. Furthermore, we performed a family-wide characterization of their biochemical properties and highlight similarities and differences in their cleavage site specificities as well as pH- and temperature-dependent activities. Detailed analysis of their sequence specificity using PICS (proteomic identification of protease cleavage sites) revealed profiles similar to human MMPs with the exception of At5-MMP; homology models of the At-MMP catalytic domains supported these results. Our results suggest that each At-MMP may be involved in different proteolytic processes during plant growth and development.
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Research Article|
December 20 2013
Family-wide characterization of matrix metalloproteinases from Arabidopsis thaliana reveals their distinct proteolytic activity and cleavage site specificity
Giada Marino;
Giada Marino
*Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden
†Umeå Plant Science Centre (UPSC), Umeå University, SE-901 87 Umeå, Sweden
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Pitter F. Huesgen;
Pitter F. Huesgen
‡Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, Canada, V6T 1Z3
§Department of Oral Biological and Medical Sciences, University of British Columbia, Vancouver, Canada, V6T 1Z3
∥Centre for Blood Research, University of British Columbia, Vancouver, Canada, V6T 1Z3
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Ulrich Eckhard;
Ulrich Eckhard
‡Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, Canada, V6T 1Z3
§Department of Oral Biological and Medical Sciences, University of British Columbia, Vancouver, Canada, V6T 1Z3
∥Centre for Blood Research, University of British Columbia, Vancouver, Canada, V6T 1Z3
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Christopher M. Overall;
Christopher M. Overall
‡Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, Canada, V6T 1Z3
§Department of Oral Biological and Medical Sciences, University of British Columbia, Vancouver, Canada, V6T 1Z3
∥Centre for Blood Research, University of British Columbia, Vancouver, Canada, V6T 1Z3
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Wolfgang P. Schröder;
Wolfgang P. Schröder
*Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden
†Umeå Plant Science Centre (UPSC), Umeå University, SE-901 87 Umeå, Sweden
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Christiane Funk
Christiane Funk
1
*Department of Chemistry, Umeå University, SE-901 87 Umeå, Sweden
†Umeå Plant Science Centre (UPSC), Umeå University, SE-901 87 Umeå, Sweden
1To whom correspondence should be addressed (email [email protected]).
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Publisher: Portland Press Ltd
Received:
February 05 2013
Revision Received:
October 16 2013
Accepted:
October 25 2013
Accepted Manuscript online:
October 25 2013
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2014 Biochemical Society
2014
Biochem J (2014) 457 (2): 335–346.
Article history
Received:
February 05 2013
Revision Received:
October 16 2013
Accepted:
October 25 2013
Accepted Manuscript online:
October 25 2013
Citation
Giada Marino, Pitter F. Huesgen, Ulrich Eckhard, Christopher M. Overall, Wolfgang P. Schröder, Christiane Funk; Family-wide characterization of matrix metalloproteinases from Arabidopsis thaliana reveals their distinct proteolytic activity and cleavage site specificity. Biochem J 15 January 2014; 457 (2): 335–346. doi: https://doi.org/10.1042/BJ20130196
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