Effect of calmodulin and its mutants on binding to Na1.2 IQ / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 71-75, 2020.
Article
in Chinese
| WPRIM
| ID: wpr-828528
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Protein Binding
/
Calmodulin
/
Calcium
/
Genetics
/
Metabolism
/
Mutation
Language:
Chinese
Journal:
Journal of Zhejiang University. Medical sciences
Year:
2020
Type:
Article
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