期刊论文

Liao, DF

英文

中国药理学报

1671-4083

2002

23

10

952-960

Project supported in part by a grant from the National Insti-tutes of Health of USA（HL-46296）and in part by a grant from the National Major Basic Research Program（973）（G2000056905）.；

本校为第一且通讯机构

AIM: To identify the specific serine/threonine residues in the C-terminal tail of thromboxane receptor α (TPα)being phosphorylated and desensitized, and various alanine mutants of these serine/threonine residues were checkedfor their ability to serve as substrates. METHODS: To facilitate the identification of the intracellular domainsinvolved in phosphorylation, glutathione S-transferase (GST)-intracellular domain fusion proteins were used assubstrates for the purified PKC, and then the cDNA of phosphorylated protein was mutagenized to locali...

目的:检测人类血栓素α受体（TPα）C端丝氨酸/苏氨酸残基的各种两氨酸诱变体作为PKC底物的能力,以确定被PKC磷酸化和脱敏的特异的丝氨酸/苏氨酸残基.方法:为了易于鉴定被磷酰化的细胞内区段,使用甘胱甘肽S-转移酶（GST）-细胞内区段融合蛋白作纯化的PKC底物,然后突变磷酰化蛋白的cDNA,以找出TPα被PKC磷酰化的主要部位,并将有组氨酸尾野生型或诱变型血栓素受体α稳定地转移到人类胚胎肾（HEK）293细胞,以研究该受体的磷酸化和脱敏.结果:仅C-端能充当纯化PKC底物.丝氨酸-331（mP4）被显示强的磷酰化,丝氨酸-324（mP1）则轻微磷酰化,丝氨酸-329（mP3）则微弱磷酰化,而其它丝氨酸/苏氨酸残基没被发...