小鼠P311蛋白与类赖氨酰氧化酶-1相互作用的鉴定

P311是利用抑制差减杂交技术从小鼠肺组织中筛选到的一个肺泡发育上游调节基因.它高水平表达于肺泡发育的相关阶段,而在慢性阻塞性肺病(chronic obstructive pulmonary disease,COPD)患者肺组织中表达水平大大下降.为深入探讨P311蛋白的作用机制,构建pGBKT7-P311诱饵表达载体,利用酵母双杂交技术,从小鼠肺组织cDNA文库中筛选出P311相互作用蛋白类赖氨酰氧化酶-1(lysyl oxidase-like 1,Loxl-1).并通过体内外免疫共沉淀及双分子荧光互补实验进行验证.为进一步研究P311蛋白的生物学功能提供新的思路。

Identification of Mouse P311 as an Interacting Protein of Lysyl Oxidase-like 1

P311 is a gene identified from mouse lung by suppression subtractive hybridization (SSH). It is highly expressed in the stages related to alveolar formation，but its expression was significantly decreased in the lungs of chronic obstructive pulmonary disease（COPD) patients. To understand the mechanism of P311 involved in lung development, a bit vector pGBKT7-P311 was used to screen a cDNA library constructed from developing mouse lungs by yeast two-hybrid system, and lysyl oxidase-like 1(Loxl-1) was identified as a partner of P311 protein. Furthermore, the interactions between P311 protein and Loxl-1 were confirmed by in vitro and in vivo coimmunoprecipitation (CoIP), and bimolecular fluorescence complementation (BiFC) experiments. This is a new approach to explore the biological function of P311 protein.