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Identification of the amino acids comprising a surface-exposed epitope within the nucleotide-binding domain of the Na+,K(+)-ATPase using a random peptide library.
Malik B
,
Jamieson GA
,
Ball WJ
.
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Monoclonal antibodies that bind native protein can generate considerable information about structure/function relationships, but identification of their epitopes can be problematic. Previously, monoclonal antibody M8-P1-A3 has been shown to bind to the catalytic (alpha) subunit of the Na+,K(+)-ATPase holoenzyme and the synthetic peptide sequence 496-HLLVMK*GAPER-506, which includes Lys 501 (K*), the major site for fluorescein-5'-isothiocyanate labeling of the Na+,K(+)-ATPase. This sequence region of alpha is proposed to comprise a portion of the enzyme's ATP binding domain (Taylor, W. R. & Green, N. W., 1989, Eur. J. Biochem. 179, 241-248). In this study we have determined M8-P1-A3's ability to recognize the alpha-subunit or homologous E1E2-ATPase proteins from different species and tissues in order to deduce the antibody's epitope. In addition the bacteriophage random peptide or "epitope" library, recently developed by Scott and Smith (1990, Science 249, 386-390) and Devlin et al. (Devlin, J. J., Panganiban, L. C., & Devlin, P. E., 1990, Science 249, 404-406), has served as a convenient technique to confirm the species-specificity mapping data and to determine the exact amino acid requirements for antibody binding. The M8-P1-A3 epitope was found to consist of the five amino acid 494-PRHLL-498 sequence stretch of alpha, with residues PRxLx being critical for antibody recognition.
Abbott,
Immunochemical and spectroscopic characterization of two fluorescein 5'-isothiocyanate labeling sites on Na+,K(+)-ATPase.
1991, Pubmed
Abbott,
Immunochemical and spectroscopic characterization of two fluorescein 5'-isothiocyanate labeling sites on Na+,K(+)-ATPase.
1991,
Pubmed
Abbott,
The epitope for the inhibitory antibody M7-PB-E9 contains Ser-646 and Asp-652 of the sheep Na+,K(+)-ATPase alpha-subunit.
1993,
Pubmed
,
Xenbase
Abbott,
The inhibitory monoclonal antibody M7-PB-E9 stabilizes E2 conformational states of Na+,K(+)-ATPase.
1992,
Pubmed
Ball,
Isolation and characterization of monoclonal antibodies to (Na+ + K+)-ATPase.
1982,
Pubmed
Ball,
Uncoupling of ATP binding to Na+,K+-ATPase from its stimulation of ouabain binding: studies of the inhibition of Na+,K+-ATPase by a monoclonal antibody.
1986,
Pubmed
Ball,
Immunochemical studies of (Na+ + K+)-ATPase using site-specific, synthetic peptide directed antibodies.
1987,
Pubmed
Ball,
Studies of the antigenic properties of sheep kidney Na+,K+-ATPase.
1982,
Pubmed
Ball,
Immunochemical comparison of cardiac glycoside-sensitive (lamb) and -insensitive (rat) kidney (Na+ + K+)-ATPase.
1986,
Pubmed
Ball,
Immunochemical evidence that the FITC-labeling site on Na+,K+-ATPase is not the ATP binding site.
1987,
Pubmed
Ball,
Preparation of antibodies to Na+,K+-ATPase and its subunits.
1988,
Pubmed
Cesareni,
Peptide display on filamentous phage capsids. A new powerful tool to study protein-ligand interaction.
1992,
Pubmed
Devlin,
Random peptide libraries: a source of specific protein binding molecules.
1990,
Pubmed
Friedman,
Determination of monoclonal antibody-induced alterations in Na+/K+-ATPase conformations using fluorescein-labeled enzyme.
1989,
Pubmed
Garnier,
Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins.
1978,
Pubmed
Geysen,
A priori delineation of a peptide which mimics a discontinuous antigenic determinant.
1986,
Pubmed
Green,
Structural modelling of P-type ion pumps.
1992,
Pubmed
Hegyvary,
Conformational changes of renal sodium plus potassium ion-transport adenosine triphosphatase labeled with fluorescein.
1981,
Pubmed
Jørgensen,
Structural basis for E1-E2 conformational transitions in Na,K-pump and Ca-pump proteins.
1988,
Pubmed
Karlish,
Characterization of conformational changes in (Na,K) ATPase labeled with fluorescein at the active site.
1980,
Pubmed
Kirley,
The amino acid sequence of the fluorescein isothiocyanate reactive site of lamb and rat kidney Na+- and K+-dependent ATPase.
1984,
Pubmed
Laemmli,
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
1970,
Pubmed
Lane,
Large-scale purification of Na,K-ATPase and its protein subunits from lamb kidney medulla.
1979,
Pubmed
Lane,
Structural studies on H+,K+-ATPase: determination of the NH2-terminal amino acid sequence and immunological cross-reactivity with Na+,K+-ATPase.
1986,
Pubmed
Maeda,
cDNA cloning and sequence determination of pig gastric (H+ + K+)-ATPase.
1988,
Pubmed
Maruyama,
Functional consequences of alterations to amino acids located in the catalytic center (isoleucine 348 to threonine 357) and nucleotide-binding domain of the Ca2+-ATPase of sarcoplasmic reticulum.
1989,
Pubmed
Parmley,
Antibody-selectable filamentous fd phage vectors: affinity purification of target genes.
1988,
Pubmed
SKOU,
The influence of some cations on an adenosine triphosphatase from peripheral nerves.
1957,
Pubmed
Scott,
Searching for peptide ligands with an epitope library.
1990,
Pubmed
Taylor,
The predicted secondary structures of the nucleotide-binding sites of six cation-transporting ATPases lead to a probable tertiary fold.
1989,
Pubmed
Tunwell,
Definition of surface-exposed epitopes on the (Ca(2+)-Mg2+)-ATPase of sarcoplasmic reticulum.
1991,
Pubmed
Tunwell,
Mapping epitopes on the (Ca(2+)-Mg2+)-ATPase of sarcoplasmic reticulum using fusion proteins.
1991,
Pubmed
Van Uem,
Determination of the epitope for the inhibitory monoclonal antibody 5-B6 on the catalytic subunit of gastric Mg(2+)-dependent H(+)-transporting and K(+)-stimulated ATPase.
1991,
Pubmed
Xu,
Nucleophilic behavior of lysine-501 of the alpha-polypeptide of sodium and potassium ion activated adenosinetriphosphatase consistent with a role in binding adenosine triphosphate.
1989,
Pubmed