Interaction between periostin and SFRP2 by co-immunoprecipitation and protein sequencing

陶熹; 王珍祥; 袁顺忠; 李世荣; 吴军

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Interaction between periostin and SFRP2 by co-immunoprecipitation and protein sequencing

更新时间：2026-01-09

- Interaction between periostin and SFRP2 by co-immunoprecipitation and protein sequencing
- Journal of Army Medical University Vol. 31, Issue 21, Pages: 2078-2080(2009)
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  CLC： R341;R394-33;R394.2
- Published：2009
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陶熹,王珍祥,袁顺忠,李世荣,吴军.免疫共沉淀-蛋白测序技术验证Periostin和SFRP2之间的相互作用[J].陆军军医大学学报(原第三军医大学学报),2009,31(21):2078-2080.

　TAO Xi,WANG Zhen-xiang,YUAN Shun-zhong,LI Shi-rong,WU Jun.Interaction between periostin and SFRP2 by co-immunoprecipitation and protein sequencing[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(21):2078-2080.
陶熹,王珍祥,袁顺忠,李世荣,吴军.免疫共沉淀-蛋白测序技术验证Periostin和SFRP2之间的相互作用[J].陆军军医大学学报(原第三军医大学学报),2009,31(21):2078-2080. DOI：

　TAO Xi,WANG Zhen-xiang,YUAN Shun-zhong,LI Shi-rong,WU Jun.Interaction between periostin and SFRP2 by co-immunoprecipitation and protein sequencing[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(21):2078-2080. DOI：

摘要

目的验证凋亡相关蛋白基因SFRP2(secreted frizzled-related protein 2)和骨母细胞特异性因子-2 Periostin（osteoblast-specific factor 2）间的相互作用。方法分别构建重组载体pCMV-Myc-SFRP2和pCMV-HA-Periostin经双酶切鉴定正确，共转染HEK293细胞，抗Myc单克隆抗体沉淀Myc-SFRP2相互作用蛋白复合物后，进行SDS-PAGE、考马斯亮蓝染色，切下目的条带，利用HPLC-Chip /MS纳流液质联用技术进行肽序列分析，经Spectrum Mill搜索Swissport数据库后鉴定蛋白质。结果经HPLC-Chip /MS纳流液质联用技术分析，免疫沉淀复合物内可检测出Periostin的表达。结论 Periostin与SFRP2间存在相互作用。

Abstract

Objective To confirm the interaction between periostin (osteoblast-specific factor 2) and secreted frizzled-related protein 2 (SFRP2) by co-immunoprecipitation and HPLC-Chip/MS techniques. Methods Expression vector encoding HA-tagged fusion protein HA-Periostin was constructed

identified and transfected into human embryo kidney 293 (HEK293) cells alone or with the expression vector pMCV-Myc-SFRP2 containing Myc-tagged fusion protein Myc-SFRP2. The interaction between SFRP2 and periostin was detected by co-immunoprecipitation with the aid of anti-Myc antibody first. Then SDS-PAGE was perform

and the blots of the protein complexes were analyzed by HPLC-Chip/MS techniques. Spectrum Mill was employed to identify the protein complexes by searching Swissport database. Results From the Myc-SFRP2 immunoprecipitation complex

periostin was identified by protein sequencing. Conclusion There is an interaction between SFRP2 and periostin.

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