NDLI: Effects of Calmodulin and Ca$^{2+}$ Channel Blockers on ω-conotoxin GVI A Binding to Crude Membranes from α$_{1B}$ Subunit (Ca$_{v}$2.2) Expressed BHK Cells and Mice Brain Lacking the α$

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Effects of Calmodulin and Ca$^{2+}$ Channel Blockers on ω-conotoxin GVI A Binding to Crude Membranes from α$_{1B}$ Subunit (Ca$_{v}$2.2) Expressed BHK Cells and Mice Brain Lacking the α$_{1B}$ Subunits

Content Provider	Springer Nature Link
Author	Wada, Tetsuyuki Imanishi, Takashi Kawaguchi, Akiri Mori, Masayuki X. Mori, Yasuo Imoto, Keiji Ichida, Seiji
Copyright Year	2005
Abstract	Characteristics for the specific binding of $^{125}$I-ω-CTX GVIA and $^{125}$I-ω-CTX MVIIC to crude membranes from BHKN101 cells expressing the α$_{1B}$ subunits of Ca$_{v}$2.2 channels and from mice brain lacking the α$_{1B}$ subunits of Ca$_{v}$2.2 channels, particularly, the effects of CaM and various Ca$^{2+}$ channel blockers on these specific bindings were investigated. Specific binding of $^{125}$I-ω-CTX GVIA to the crude membranes from BHKN101 cells was observed, but not from control BHK6 cells. ω-CTX GVIA, ω-CTX MVIIC and ω-CTX SVIB inhibited the specific binding of $^{125}$I-ω-CTX GVIA to crude membranes from BHKN101 cells, and the IC$_{50}$ values for ω-CTXGVIA, ω-CTX MVIIC and ω-CTX SVIB were 0.07, 8.5 and 1.7 nM, respectively. However, ω-agatoxin IVA and calciseptine at concentrations of 10$^{−9}$–10$^{−6}$ M did not inhibit specific binding. Specific binding was also about 80% inhibited by 20 μg protein/ml CaM. The amount of $^{125}$I-ω-CTX GVIA (30 pM) specifically bound to membranes from brain of knockout mice lacking α$_{1B}$ subunits of Ca$_{v}$2.2 channels was about 30% of that to the crude membranes from brain of wild-type. On the other hand, specific binding of $^{125}$I-ω-CTX MVIIC (200 pM) was observed on the crude membranes of both BHKN101 and control BHK6 cells. The specific binding of $^{125}$I-ω-CTX MVIIC (200 pM) was not inhibited by ω-CTX GVIA and ω-CTX SVIB, and also ω-Aga IVA and calciseptine at concentrations of 10$^{−9}$–10$^{−7}$ M, although specific binding was almost completely dose dependently inhibited by non-radiolabeled ω-CTX MVIIC (IC$_{50}$ value was about 0.1 nM). 20 μg protein/ml CaM did not inhibit specific binding. Therefore, these results suggest that BHKN101 cells have a typical Ca$_{v}$2.2 channels which are also inhibited by CaM and have not specific binding sites for ω-CTX MVIIC, although ω-CTX MVIIC is a blocker for both Ca$_{v}$2.1 (α$_{1A; }$P/Q-type) and Ca$_{v}$2.2 channels.
Starting Page	1045
Ending Page	1054
Page Count	10
File Format	PDF
ISSN	03643190
Journal	Neurochemical Research
Volume Number	30
Issue Number	8
e-ISSN	15736903
Language	English
Publisher	Kluwer Academic Publishers-Plenum Publishers
Publisher Date	2005-01-01
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	BHK cells expressed α$_{1B }$(Ca$_{v}$ 2.2) subunit Ca$_{v}$2.2 (N-type) channels Ca$^{2+}$ channels blockers calmodulin KO mice of α$_{1B }$(Ca$_{v}$2.2)subunit Omega-conotoxin GVIA Omega-conotoxin MVIIC Neurosciences Neurology Biochemistry
Content Type	Text
Resource Type	Article
Subject	Medicine Biochemistry Cellular and Molecular Neuroscience

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Effects of Calmodulin and Ca$^{2+}$ Channel Blockers on ω-conotoxin GVI A Binding to Crude Membranes from α$_{1B}$ Subunit (Ca$_{v}$2.2) Expressed BHK Cells and Mice Brain Lacking the α$_{1B}$ Subunits

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