Effect of calmodulin and its mutants on binding to Na1.2 IQ.
- Author:
Yujun WAN
1
;
Junyan LIU
2
;
Yuting WANG
2
;
Xiaoyu CHENG
1
;
Sha SHA
1
;
Wanying JIA
2
;
Delin HU
1
;
Xinyu LI
1
;
Feng GUO
2
Author Information
1. Department of Clinical Medicine, China Medical University, Shenyang 110122, China.
2. Department of Pharmaceutical Toxicology, School of Pharmacy, China Medical University, Shenyang 110122, China.
- Publication Type:Journal Article
- MeSH:
Calcium;
metabolism;
Calmodulin;
genetics;
metabolism;
Mutation;
NAV1.2 Voltage-Gated Sodium Channel;
metabolism;
Protein Binding;
genetics
- From:
Journal of Zhejiang University. Medical sciences
2020;49(1):71-75
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of calmodulin (CaM) and its mutants on binding to voltage-gated Na channel isoleucine-glutamine domain (Na1.2 IQ).
METHODS:The cDNA of Na1.2 IQ was constructed by PCR technique, CaM mutants CaM, CaM and CaM were constructed with Quickchange site-directed mutagenesis kit (QIAGEN). The binding of Na1.2 IQ to CaM and CaM mutants under calcium and calcium free conditions were detected by pull-down assay.
RESULTS:Na1.2 IQ and CaM were bound to each other at different calcium concentrations, while GST alone did not bind to CaM. The binding affinity of CaM and Na1.2 IQ at [Ca]-free was greater than that at 100 nmol/L [Ca] ( < 0.05). In the absence of calcium, the binding amount of CaM wild-type to Na1.2 IQ was greater than that of its mutant, and the binding affinity of CaM to Na1.2 IQ was the weakest among the three mutants ( < 0.05).
CONCLUSIONS:The binding ability of CaM and CaM mutants to Na1.2 IQ is Ca-dependent. This study has revealed a new mechanism of Na1.2 regulated by CaM, which would be useful for the study of ion channel related diseases.
