Immunochemical evidence that the FITC-labeling site on Na +,K +-ATPase is not the ATP binding site

The covalent labeling of the α subunit of lamb kidney Na +,K +-ATPase by fluorescein 5′-isothiocyanate at Lys-501 has generally been assumed to occur at the ATP binding site. We have found that the peptide sequence 496HLLVMKGAPER 506 serves as the antigenic determinant for monoclonal antibody M8-P1-...

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Bibliographic Details
Published inBiochemical and biophysical research communications Vol. 148; no. 1; pp. 246 - 253
Main Authors Ball, W.J., Friedman, M.L.
Format Journal Article
LanguageEnglish
Published San Diego, CA Elsevier Inc 14.10.1987
Elsevier
Subjects
Adenosine Triphosphate - metabolism
Amino Acid Sequence
Animals
Antibodies, Monoclonal
Applied sciences
Binding Sites
Binding, Competitive
Epitopes - analysis
Exact sciences and technology
Fluorescein-5-isothiocyanate
Fluoresceins - metabolism
Fluorescent Dyes - metabolism
Kidney - enzymology
Kinetics
Macromolecular Substances
Other techniques and industries
Protein Binding
Sheep
Sodium-Potassium-Exchanging ATPase - immunology
Sodium-Potassium-Exchanging ATPase - metabolism
Thiocyanates - metabolism
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Summary:The covalent labeling of the α subunit of lamb kidney Na +,K +-ATPase by fluorescein 5′-isothiocyanate at Lys-501 has generally been assumed to occur at the ATP binding site. We have found that the peptide sequence 496HLLVMKGAPER 506 serves as the antigenic determinant for monoclonal antibody M8-P1-A3. This antibody binds to both native and FITC-labeled enzyme and while this epitope undergoes ligand-induced changes these changes are not involved in either enzyme function or the E 1 ⇄ E 2 conformational changes monitored by FITC-fluorescence intensity.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:0006-291X
1090-2104
DOI:10.1016/0006-291X(87)91102-8