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The Journal of Neuroscience, January 15, 1998, 18(2):641-647

Evidence for a 95 kDa Short Form of the _1A Subunit Associated with the -Conotoxin MVIIC Receptor of the P/Q-type Ca²⁺ Channels

Victoria E. S. Scott¹, Ricardo Felix¹, Jyothi Arikkath^{1, 2}, and Kevin P. Campbell¹

¹ Howard Hughes Medical Institute, Departments of Physiology and Biophysics, Neurology, and ² Program in Molecular Biology, University of Iowa College of Medicine, Iowa City, Iowa 52242

Neuronal voltage-dependent Ca²⁺ channels have been isolated previously and shown to contain a primary ₁ pore-forming subunit as well as auxiliary ₂ and  subunits, in addition to an uncharacterized 95 kDa protein. In the present study, using multiple approaches, we have extensively characterized the molecular structure of the 95 kDa protein. Separation of the P/Q- and N-type neuronal Ca²⁺ channels showed that the 95 kDa protein is associated exclusively with the -Conotoxin MVIIC receptor of the P/Q-type channels. Analysis of purified synaptic plasma membranes and the isolated P/Q-type channels, using _1A-specific antibodies, suggested a structural relationship between the _1A subunit and the 95 kDa protein. This finding was supported by protein-protein interaction data, which revealed that the  subunit can associate with the 95 kDa protein in addition to the _1A subunit. Changes in electrophoretic mobility after enzymatic treatment with Endo F indicated that the 95 kDa protein is glycosylated. Furthermore, microsequencing of the 95 kDa protein yielded 13 peptide sequences, all of which are present in the first half of the _1A subunit up to amino acid 829 of the cytoplasmic linker between repeats II and III. Taken together, our results strongly suggest that the 95 kDa glycoprotein associated with the P/Q-type Ca²⁺ channels is a short form of the _1A subunit.

Key words: P/Q-type Ca²⁺ channels; 95 kDa subunit; _1A subunit; N-type Ca²⁺ channel; subunit; -Conotoxin MVIIC; -Conotoxin GVIA

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Copyright © 1998 Society for Neuroscience  0270-6474/98/182641-07$05.00/0

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