[KCNE2 protein S98 phosphorylation in heart of old SHR rats detected by point mutagenesis].
To investigate the phosphorylation of KCNE2 protein in heart of old SHR rats.



The membrane proteins from ventricular myocardium of old SHR were extracted, treated with or without alkaline phosphatase and tested binding with Ab2 (an anti-KCNE2 polyclonal antibody) by Western blot. A KCNE2 fusion protein with c-myc was obtained from in vitro translation system and treated with or without alkaline phosphatase. A series of mono- and double-point mutated fusion KCNE2 proteins with c-myc were obtained from an in vitro translation system, and Western blots with Ab2 or anti-myc antibody were performed.



After alkaline phosphatase treatment, Ab2 significantly attenuated its binding with KCNE2. In vitro translation system confirmed that after alkaline phosphatase treatment, Ab2 weakened binding ability to KCNE2 while binding to c-myc was not changed. Point mutation experiments showed that serine residue in position 98 of KCNE2 proteins might be phosphorylated.



KCNE2 protein in heart of old SHR rats is phosphorylated and this phosphorylation takes place in serine residue of position 98.
Wang
Hui-lian
HL
Department of Vascular Surgery, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, 310003, China. hlianw@yahoo.com.cn
Lu
She-min
SM
Ning
Qi-lan
QL
D000375:Aging; D000818:Animals; D015153:Blotting, Western; D006973:Hypertension / Q000235:genetics / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D010766:Phosphorylation; D017354:Point Mutation; D024642:Potassium Channels, Voltage-Gated / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011918:Rats, Inbred SHR; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
C489621:KCNE2 protein, rat; D024642:Potassium Channels, Voltage-Gated; D016271:Proto-Oncogene Proteins c-myc; D011993:Recombinant Fusion Proteins