Physicochemical characterization of the alpha-peptide of the sodium channel from rat brain.
The alpha-peptide of the rat brain sodium channel of apparent molecular weight 260K has been purified to homogeneity in order to determine its structural and chemical properties. By negative-stain electron microscopy, the molecule morphology of the solubilized channel protein appears as a stack of disks or rouleaux whose dimensions are 40 A X 200 A. Measurement of the secondary structure by circular dichroism shows that the alpha-peptide is a conformationally flexible polypeptide that contains mostly beta-sheet and random-coil in mixed detergent-phospholipid micelles and folds into a conformation that has approximately 65% alpha-helix after reconstitution into phosphatidylcholine vesicles. Preparative polyacrylamide gel electrophoresis was used to obtain a chemically homogeneous peptide to analyze the amino acid and carbohydrate composition. The amino acid composition shows a reasonably high content of acidic amino acids with no striking excess of hydrophobic amino acids, while carbohydrate analyses show that carbohydrate is 31% by weight of the protein with sialic acid representing over 50% of the total carbohydrates. The high alpha-helical content, the amino acid composition, and the large carbohydrate mass are similar to those of the eel electroplax sodium channel and appear to be general features of the sodium channels which have been analyzed structurally and chemically to date.
L W
LW
Elmer
B J
BJ
O'Brien
T J
TJ
Nutter
K J
KJ
Angelides
D000818:Animals; D001923:Brain Chemistry; D002462:Cell Membrane / Q000378:metabolism; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D007473:Ion Channels / Q000032:analysis* / Q000378:metabolism; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000302:isolation & purification* / Q000378:metabolism; D011487:Protein Conformation; D051381:Rats; D012530:Saxitoxin / Q000378:metabolism; D012964:Sodium / Q000378:metabolism*; D015222:Sodium Channels
D007473:Ion Channels; D009419:Nerve Tissue Proteins; D015222:Sodium Channels; D012530:Saxitoxin; D012964:Sodium