The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis
Abstract Background Connective tissue growth factor (CTGF)-induced angiogenesis is a crucial factor in rheumatoid arthritis (RA), but CTGF-interacting protein and related molecular mechanism of their interaction have not been fully elucidated. Methods CTGF-interacting proteins were identified throug...
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| Format: | Article |
| Language: | English |
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BMC
2021-10-01
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| Series: | Arthritis Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13075-021-02656-y |
| _version_ | 1819003823762440192 |
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| author | Guoyu Yin Chenglin Yang Gan Wu Xinxin Yu Qingqing Tian Daoxing Chen Ben Cao Lin Zhao Nannan Xu Shengwei Jin Wei Zhang Jianguang Wang |
| author_facet | Guoyu Yin Chenglin Yang Gan Wu Xinxin Yu Qingqing Tian Daoxing Chen Ben Cao Lin Zhao Nannan Xu Shengwei Jin Wei Zhang Jianguang Wang |
| author_sort | Guoyu Yin |
| collection | DOAJ |
| description | Abstract Background Connective tissue growth factor (CTGF)-induced angiogenesis is a crucial factor in rheumatoid arthritis (RA), but CTGF-interacting protein and related molecular mechanism of their interaction have not been fully elucidated. Methods CTGF-interacting proteins were identified through the LC-MS/MS analysis of the Co-IP products from fibroblast-like synoviocyte (FLS) lysates, and the interaction between CTGF and annexin A2 (ANXA2) was further confirmed through Co-IP and BiFC assay. The binding domain, mutant, mechanism, and angiogenesis function were assessed by homology modeling, molecular docking, MTT, cell scratch, tube formation, and chick chorioallantoic membrane (CAM) assays. Additionally, severe combined immunodeficiency (SCID) mouse co-implantation model was constructed to confirm the effect of ANXA2/CTGF-TSP1 in the process of RA in vivo. Results ANXA2 was identified and verified as an interaction partner of CTGF for the first time by Co-IP and LC-MS/MS analysis. Co-localization of CTGF and ANXA2 was observed in RA-FLS, and direct interaction of the TSP-1 domain of CTGF and ANXA2 was determined in HEK293T cells. The spatial conformation and stable combination of the ANXA2/CTGF-TSP1 complex were assessed by homology modeling in the biomimetic environment. The function of the ANXA2/CTGF-TSP1 complex was proved on promoting FLS proliferation, migration, and angiogenesis in vitro and deteriorating FLS invasion and joint damage in SCID mice. Conclusions TSP-1 is the essential domain in CTGF/ANXA2 interaction and contributes to FLS migration and pannus formation, inducing the process of RA. |
| first_indexed | 2024-12-20T23:27:08Z |
| format | Article |
| id | doaj.art-73b0902edbf448b997f379ea775d2af5 |
| institution | Directory Open Access Journal |
| issn | 1478-6362 |
| language | English |
| last_indexed | 2024-12-20T23:27:08Z |
| publishDate | 2021-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Arthritis Research & Therapy |
| spelling | doaj.art-73b0902edbf448b997f379ea775d2af52022-12-21T19:23:22ZengBMCArthritis Research & Therapy1478-63622021-10-0123111310.1186/s13075-021-02656-yThe protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritisGuoyu Yin0Chenglin Yang1Gan Wu2Xinxin Yu3Qingqing Tian4Daoxing Chen5Ben Cao6Lin Zhao7Nannan Xu8Shengwei Jin9Wei Zhang10Jianguang Wang11Department of Anesthesia and Critical Care, School of the Second Clinical Medical Sciences, Wenzhou Medical UniversityQiushi Academy for Advanced Studies, Zhejiang UniversityDepartment of Anesthesia and Critical Care, School of the Second Clinical Medical Sciences, Wenzhou Medical UniversityDepartment of Biochemistry, School of Basic Medical Sciences, Wenzhou Medical UniversityDepartment of Biochemistry, School of Basic Medical Sciences, Wenzhou Medical UniversityDepartment of Medicinal Chemistry, School of Pharmaceutical Sciences, Wenzhou Medical UniversityDepartment of Biochemistry, School of Basic Medical Sciences, Wenzhou Medical UniversityDepartment of Biochemistry, School of Basic Medical Sciences, Wenzhou Medical UniversityDepartment of Biochemistry, School of Basic Medical Sciences, Wenzhou Medical UniversityDepartment of Anesthesia and Critical Care, School of the Second Clinical Medical Sciences, Wenzhou Medical UniversityDepartment of Biochemistry, School of Basic Medical Sciences, Wenzhou Medical UniversityDepartment of Anesthesia and Critical Care, School of the Second Clinical Medical Sciences, Wenzhou Medical UniversityAbstract Background Connective tissue growth factor (CTGF)-induced angiogenesis is a crucial factor in rheumatoid arthritis (RA), but CTGF-interacting protein and related molecular mechanism of their interaction have not been fully elucidated. Methods CTGF-interacting proteins were identified through the LC-MS/MS analysis of the Co-IP products from fibroblast-like synoviocyte (FLS) lysates, and the interaction between CTGF and annexin A2 (ANXA2) was further confirmed through Co-IP and BiFC assay. The binding domain, mutant, mechanism, and angiogenesis function were assessed by homology modeling, molecular docking, MTT, cell scratch, tube formation, and chick chorioallantoic membrane (CAM) assays. Additionally, severe combined immunodeficiency (SCID) mouse co-implantation model was constructed to confirm the effect of ANXA2/CTGF-TSP1 in the process of RA in vivo. Results ANXA2 was identified and verified as an interaction partner of CTGF for the first time by Co-IP and LC-MS/MS analysis. Co-localization of CTGF and ANXA2 was observed in RA-FLS, and direct interaction of the TSP-1 domain of CTGF and ANXA2 was determined in HEK293T cells. The spatial conformation and stable combination of the ANXA2/CTGF-TSP1 complex were assessed by homology modeling in the biomimetic environment. The function of the ANXA2/CTGF-TSP1 complex was proved on promoting FLS proliferation, migration, and angiogenesis in vitro and deteriorating FLS invasion and joint damage in SCID mice. Conclusions TSP-1 is the essential domain in CTGF/ANXA2 interaction and contributes to FLS migration and pannus formation, inducing the process of RA.https://doi.org/10.1186/s13075-021-02656-yRheumatoid arthritis (RA)Pannus formationAnnexin A2 (ANXA2)Connective tissue growth factor (CTGF) |
| spellingShingle | Guoyu Yin Chenglin Yang Gan Wu Xinxin Yu Qingqing Tian Daoxing Chen Ben Cao Lin Zhao Nannan Xu Shengwei Jin Wei Zhang Jianguang Wang The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis Arthritis Research & Therapy Rheumatoid arthritis (RA) Pannus formation Annexin A2 (ANXA2) Connective tissue growth factor (CTGF) |
| title | The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis |
| title_full | The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis |
| title_fullStr | The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis |
| title_full_unstemmed | The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis |
| title_short | The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis |
| title_sort | protein protein interaction between connective tissue growth factor and annexin a2 is relevant to pannus formation in rheumatoid arthritis |
| topic | Rheumatoid arthritis (RA) Pannus formation Annexin A2 (ANXA2) Connective tissue growth factor (CTGF) |
| url | https://doi.org/10.1186/s13075-021-02656-y |
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