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http://dx.doi.org/10.4014/jmb.1501.01066

Asn-Linked Glycosylation Contributes to Surface Expression and Voltage-Dependent Gating of Cav1.2 Ca2+ Channel  

Park, Hyun-Jee (Department of Life Science, Sogang University)
Min, Se-Hong (Department of Life Science, Sogang University)
Won, Yu-Jin (Department of Life Science, Sogang University)
Lee, Jung-Ha (Department of Life Science, Sogang University)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.8, 2015 , pp. 1371-1379 More about this Journal
Abstract
The Cav1.2 Ca2+ channel is essential for cardiac and smooth muscle contractility and many physiological functions. We mutated single, double, and quadruple sites of the four potential Asn (N)-glycosylation sites in the rabbit Cav1.2 into Gln (Q) to explore the effects of Nglycosylation. When a single mutant (N124Q, N299Q, N1359Q, or N1410Q) or Cav1.2/WT was expressed in Xenopus oocytes, the biophysical properties of single mutants were not significantly different from Cav1.2/WT. In comparison, the double mutant N124,299Q showed a positive shift in voltage-dependent gating. Furthermore, the quadruple mutant (QM; N124,299,1359,1410Q) showed a positive shift in voltage-dependent gating as well as a reduction of current. We tagged EGFP to the QM, double mutants, and Cav1.2/WT to chase the mechanisms underlying the reduced currents of QM. The surface fluorescence intensity of QM was weaker than that of Cav1.2/WT, suggesting that the reduced current of QM arises from its lower surface expression than Cav1.2/WT. Tunicamycin treatment of oocytes expressing Cav1.2/WT mimicked the effects of the quadruple mutations. These findings suggest that Nglycosylation contributes to the surface expression and voltage-dependent gating of Cav1.2.
Keywords
N-glycosylation; Cav1.2 Ca2+ channel; point mutation; Xenopus oocyte; voltage clamping;
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