免疫共沉淀结合微流控芯片技术筛选GBLP相互作用蛋白

Screening of GBLP-interacting Proteins with Co-immunoprecipitation and an Integrated Microfluidic System

为了进一步阐明G蛋白β亚基2样1蛋白(guanine nucleotide-binding protein subunit beta 2-like 1,GBLP)在普萘洛尔(propranolol,PRO)生物学效应发生机制中的作用,采用免疫共沉淀法结合液相色谱-芯片-离子阱串联质谱(HPLC-CHIP-IT-MS/MS)系统筛选并鉴定人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)中的GBLP相互作用蛋白.结果表明,有7种蛋白与GBLP存在相互作用,分别为酪蛋白α-S1、酪蛋白α-S2、β酪蛋白、桥粒芯蛋白1前体、α-烯醇化酶、果糖二磷酸醛缩酶C、硫氧还原蛋白过氧化物酶2.这些蛋白的生物信息学检索结果提示,GBLP可能参与了细胞的能量代谢调节和抗氧化机制,与普萘洛尔的生物学效应发生机制密切相关.

To further elucidate the biological characteristics and the roles of guanine nucleotide-binding protein subunit beta 2-like 1 （GBLP） in response to propranolol （PRO）, co-immunoprecipitation and HPLC-CHIP-ITMS/MS were used to screen and identity the GBLP interacting proteins in human umbilical vein endothelial ceils. Seven proteins were shown to interact with GBLP, including casein α-S1, casein α-S2, β casein, desmoglein 1 precursor, α-enolase, fructose-bisphosphate aldolase C and peroxiredoxin 2. These proteins actively involved in energy metabolism of the cell, suggesting that GBLP might participated in the regulation of antioxidation processes, which correlated with the known biological functions of PRO.