Purification of putative Ca2+ channel protein from rabbit skeletal muscle. Determination of the amino-terminal sequence.
A putative Ca2+ channel protein was purified from rabbit skeletal muscle transverse tubules with the combined use of lectin affinity chromatography and ion-exchange chromatography, followed by sucrose density gradient centrifugation. The major component of the purified preparation detected by sodium dodecyl sulfate-gel electrophoresis was a protein of 150 kDa when reduced with 20 mM dithiothreitol and a 191-kDa protein when treated with 20 mM N-ethylmaleimide. Therefore, this protein appears to be identical with the alpha subunit previously described (Curtis, B. M., and Catterall, W. A. (1984) Biochemistry 23, 2113-2118). This protein was purified by preparative sodium dodecyl sulfate-gel electrophoresis, followed by electroelution and/or electroblotting, and its amino acid composition and NH2-terminal sequence were determined. The NH2-terminal sequence is: NH2-Glu-Pro-Phe-Pro-Ser-Ala-Val-X-Ile-Lys-Ser-X-Val-X-Lys-Met-Gln-.
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels; D007473:Ion Channels / Q000378:metabolism*; D008565:Membrane Proteins / Q000302:isolation & purification; D008970:Molecular Weight; D009124:Muscle Proteins / Q000302:isolation & purification*; D009132:Muscles / Q000378:metabolism*; D011817:Rabbits; D011978:Receptors, Nicotinic / Q000302:isolation & purification*; D012995:Solubility
D015220:Calcium Channels; D007473:Ion Channels; D008565:Membrane Proteins; D009124:Muscle Proteins; D011978:Receptors, Nicotinic; D002118:Calcium