Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids
ObjectivesTo explore posttranslational modifications (PTMs), including proteolytic activation, multimerization, complex formation and citrullination of gelatinases, in particular of gelatinase B/MMP-9, and to detect in gelatin-Sepharose affinity-purified synovial fluids, the presence of specific MMP...
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Frontiers Media S.A.
2021
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oai:doaj.org-article:fcdb7e2221a94516bc4d928080b5e8a72021-12-01T13:37:39ZProteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids1664-322410.3389/fimmu.2021.763832https://doaj.org/article/fcdb7e2221a94516bc4d928080b5e8a72021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fimmu.2021.763832/fullhttps://doaj.org/toc/1664-3224ObjectivesTo explore posttranslational modifications (PTMs), including proteolytic activation, multimerization, complex formation and citrullination of gelatinases, in particular of gelatinase B/MMP-9, and to detect in gelatin-Sepharose affinity-purified synovial fluids, the presence of specific MMP proteoforms in relation to arthritis.MethodsLatent, activated, complexed and truncated gelatinase-A/MMP-2 and gelatinase B/MMP-9 proteoforms were detected with the use of zymography analysis to compare specific levels, with substrate conversion assays, to test net proteolytic activities and by Western blot analysis to decipher truncation variants. Citrullination was detected with enhanced sensitivity, by the use of a new monoclonal antibody against modified citrullines.ResultsAll MMP-9 and MMP-2 proteoforms were identified in archival synovial fluids with the use of zymography analysis and the levels of MMP-9 versus MMP-2 were studied in various arthritic diseases, including rheumatoid arthritis (RA). Secondly, we resolved misinterpretations of MMP-9 levels versus proteolytic activities. Thirdly, a citrullinated, truncated proteoform of MMP-9 was discovered in archival RA synovial fluid samples and its presence was corroborated as citrullinated hemopexin-less MMP-9 in a small prospective RA sample cohort.ConclusionSynovial fluids from rheumatoid arthritis contain high levels of MMP-9, including its truncated and citrullinated proteoform. The combination of MMP-9 as analyte and its PTM by citrullination could be of clinical interest, especially in the field of arthritic diseases.Bernard GrilletKaren YuEstefania Ugarte-BerzalRik JanssensRafaela Vaz Sousa PereiraLise BoonErik MartensNele BerghmansIsabelle RonsseIlse Van AelstPierre FitenRené ConingsJennifer VandoorenPatrick VerschuerenJo Van DammePaul ProostGhislain OpdenakkerFrontiers Media S.A.articlematrix metalloproteinasearthritisproteoformproteolysiscitrullinationsynovial fluidImmunologic diseases. AllergyRC581-607ENFrontiers in Immunology, Vol 12 (2021) |
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matrix metalloproteinase arthritis proteoform proteolysis citrullination synovial fluid Immunologic diseases. Allergy RC581-607 |
| spellingShingle |
matrix metalloproteinase arthritis proteoform proteolysis citrullination synovial fluid Immunologic diseases. Allergy RC581-607 Bernard Grillet Karen Yu Estefania Ugarte-Berzal Rik Janssens Rafaela Vaz Sousa Pereira Lise Boon Erik Martens Nele Berghmans Isabelle Ronsse Ilse Van Aelst Pierre Fiten René Conings Jennifer Vandooren Patrick Verschueren Jo Van Damme Paul Proost Ghislain Opdenakker Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids |
| description |
ObjectivesTo explore posttranslational modifications (PTMs), including proteolytic activation, multimerization, complex formation and citrullination of gelatinases, in particular of gelatinase B/MMP-9, and to detect in gelatin-Sepharose affinity-purified synovial fluids, the presence of specific MMP proteoforms in relation to arthritis.MethodsLatent, activated, complexed and truncated gelatinase-A/MMP-2 and gelatinase B/MMP-9 proteoforms were detected with the use of zymography analysis to compare specific levels, with substrate conversion assays, to test net proteolytic activities and by Western blot analysis to decipher truncation variants. Citrullination was detected with enhanced sensitivity, by the use of a new monoclonal antibody against modified citrullines.ResultsAll MMP-9 and MMP-2 proteoforms were identified in archival synovial fluids with the use of zymography analysis and the levels of MMP-9 versus MMP-2 were studied in various arthritic diseases, including rheumatoid arthritis (RA). Secondly, we resolved misinterpretations of MMP-9 levels versus proteolytic activities. Thirdly, a citrullinated, truncated proteoform of MMP-9 was discovered in archival RA synovial fluid samples and its presence was corroborated as citrullinated hemopexin-less MMP-9 in a small prospective RA sample cohort.ConclusionSynovial fluids from rheumatoid arthritis contain high levels of MMP-9, including its truncated and citrullinated proteoform. The combination of MMP-9 as analyte and its PTM by citrullination could be of clinical interest, especially in the field of arthritic diseases. |
| format |
article |
| author |
Bernard Grillet Karen Yu Estefania Ugarte-Berzal Rik Janssens Rafaela Vaz Sousa Pereira Lise Boon Erik Martens Nele Berghmans Isabelle Ronsse Ilse Van Aelst Pierre Fiten René Conings Jennifer Vandooren Patrick Verschueren Jo Van Damme Paul Proost Ghislain Opdenakker |
| author_facet |
Bernard Grillet Karen Yu Estefania Ugarte-Berzal Rik Janssens Rafaela Vaz Sousa Pereira Lise Boon Erik Martens Nele Berghmans Isabelle Ronsse Ilse Van Aelst Pierre Fiten René Conings Jennifer Vandooren Patrick Verschueren Jo Van Damme Paul Proost Ghislain Opdenakker |
| author_sort |
Bernard Grillet |
| title |
Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids |
| title_short |
Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids |
| title_full |
Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids |
| title_fullStr |
Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids |
| title_full_unstemmed |
Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids |
| title_sort |
proteoform analysis of matrix metalloproteinase-9/gelatinase b and discovery of its citrullination in rheumatoid arthritis synovial fluids |
| publisher |
Frontiers Media S.A. |
| publishDate |
2021 |
| url |
https://doaj.org/article/fcdb7e2221a94516bc4d928080b5e8a7 |
| work_keys_str_mv |
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