Location of disulfide bonds in the Na+, K(+)-ATPase alpha subunit.
For localizing S-S bonds in the pig kidney Na+,K(+)-ATPase alpha subunit, cystine-containing peptides (V-1, VII-1, and VII-2), obtained in our previous study from the enzyme's tryptic digest, were analysed. Chemical modification of the cystine-containing peptides performed at cysteine residues involved successive alkylation, first with radioactive iodoacetic acid and then with ABD-F in the absence and presence of a reducing agent, respectively. Cysteinyl peptides were isolated by HPLC, their amino acid sequences determined, and two disulfide bonds: Cys452-Cys456 and Cys511-Cys549 were localized by identification of fluorescent cysteine residues.
Ovchinnikov and Shemyakin Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow.
N M
NM
Gevondyan
V S
VS
Gevondyan
N N
NN
Modyanov
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003553:Cystine / Q000032:analysis; D004220:Disulfides / Q000032:analysis*; D007679:Kidney Medulla / Q000201:enzymology; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry*; D013552:Swine; D014357:Trypsin
D004220:Disulfides; D046911:Macromolecular Substances; D010446:Peptide Fragments; D003553:Cystine; D014357:Trypsin; D000254:Sodium-Potassium-Exchanging ATPase