The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis.
The protein-protein interaction between connective tissue growth factor and annexin A2 is relevant to pannus formation in rheumatoid arthritis.
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DOI：
10.1186/s13075-021-02656-y
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发表时间：
2021-10-26
影响因子：
4.9
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Yin G;Yang C;Wu G;Yu X;Tian Q;Chen D;Cao B;Zhao L;Xu N;Jin S;Zhang W;Wang J
结缔组织生长因子（CTGF）诱导的血管生成是类风湿性关节炎（RA）的关键因素，但CTGF相互作用蛋白及其相互作用的分子机制尚未完全阐明。通过LC-MS/MS分析成纤维细胞样滑膜细胞（FLS）裂解物的Co-IP产物来鉴定CTGF相互作用蛋白，并且通过Co-IP和BiFC分析进一步证实CTGF与膜联蛋白A2（ANXA 2）之间的相互作用。通过同源性建模、分子对接、MTT、细胞划痕、管形成和鸡胚绒毛尿囊膜（CAM）测定来评估结合结构域、突变体、机制和血管生成功能。此外，本研究还建立了严重联合免疫缺陷（severe combined immunodeficiency，SCID）小鼠共植入模型，以证实ANXA 2/CTGF-TSP 1在RA发病过程中的作用。通过Co-IP和LC-MS/MS分析，ANXA 2首次被鉴定并验证为CTGF的相互作用伙伴。在RA-FLS中观察到CTGF和ANXA 2的共定位，并且在HEK 293 T细胞中确定CTGF和ANXA 2的TSP-1结构域的直接相互作用。在仿生环境中通过同源模建评估ANXA 2/CTGF-TSP 1复合物的空间构象和稳定结合。ANXA 2/CTGF-TSP 1复合物在体外可促进FLS增殖、迁移和血管生成，在SCID小鼠中可减轻FLS侵袭和关节损伤。TSP-1是CTGF/ANXA 2相互作用的关键结构域，参与FLS迁移和血管翳形成，诱导RA的发生。在线版本包含补充材料，可通过10.1186/s13075-021-02656-y获得。
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. 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. 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. TSP-1 is the essential domain in CTGF/ANXA2 interaction and contributes to FLS migration and pannus formation, inducing the process of RA. The online version contains supplementary material available at 10.1186/s13075-021-02656-y.
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影响因子：
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作者：
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通讯作者：
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