Inactivation of the Na,K‐ATPase by modification of Lys‐501 with 4‐acetamido‐4′‐isothiocyanatostilbene‐2,2′‐disulfonic acid (SITS)
- 7 December 1992
- journal article
- Published by Wiley in FEBS Letters
- Vol. 314 (1) , 97-100
- https://doi.org/10.1016/0014-5793(92)81470-7
Abstract
The sodium pump or Na,K‐ATPase, maintains the Na+ and K+ gradients across eukaryotic cell membranes at the expense of ATP. Incubation of purified canine renal Na,K‐ATPase with 4‐acetamido‐4′‐isothiocyanatostilbene‐2,2′‐disulfonic acid (SITS) inhibited the ATPase activity. Both the labeling of the protein and the loss or ATPase activity were prevented by co‐incubation with ADP (acting as an ATP analog) or KCI. Only the α‐subunit was labeled by SITS. The α‐subunit from the inhibited enzyme was extensively digested with trypsin, and SITS‐labeled peptides were purified by reverse‐phase HPLC and sequenced. The amino acid sequence determined, His‐Leu‐Leu‐Val‐Met‐X‐Gly‐Ala‐Pro‐Glu, indicated that SITS modifies Lys‐501 (X) on the α‐subunit of Na,K‐ATPase.Keywords
This publication has 14 references indexed in Scilit:
- Chemical modification as an approach to elucidation of sodium pump structure-function relationsAmerican Journal of Physiology-Cell Physiology, 1990
- Inhibition and derivatization of the renal sodium-potassium-ATPase by dihydro-4,4'-diisothiocyanatostilbene-2,2'-disulfonateBiochemistry, 1988
- Mutation of aspartic acid-351, lysine-352, and lysine-515 alters the Ca2+ transport activity of the Ca2+-ATPase expressed in COS-1 cells.Proceedings of the National Academy of Sciences, 1988
- [9] Preparation of antibodies to Na+,K+-ATPase and its subunitsPublished by Elsevier ,1988
- Immunochemical evidence that the FITC-labeling site on Na+,K+-ATPase is not the ATP binding siteBiochemical and Biophysical Research Communications, 1987
- Amino-acid sequence of the catalytic subunit of the (Na+ + K+)ATPase deduced from a complementary DNANature, 1985
- The amino acid sequence of the fluorescein isothiocyanate reactive site of lamb and rat kidney Na+- and K+-dependent ATPaseBiochemical and Biophysical Research Communications, 1984
- The effect of anion channel blockers on enzymatic activity of Na+/K+‐ATPase and the electrogenic Na+/K+ pumpFEBS Letters, 1984
- Effects of the stilbene derivatives SITS and DIDS on intestinal ATPase activitiesPharmacological Research Communications, 1984
- Anion transport across the erythrocyte membrane, in situ proteolysis of band 3 protein, and cross-linking of proteolytic fragments by 4,4′-diisothiocyano dihydrostilbene-2,2′-disulfonateBiochimica et Biophysica Acta (BBA) - Biomembranes, 1979