Modification of chloride ion transport in human erythrocyte ghost membranes by photoaffinity labeling reagents based on the structure of 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB). 1995

B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
Department of Chemistry, Connecticut College, New London 06320, USA.

Using human red blood cell ghost membranes, we have evaluated 5-nitro-2-[N-3-(4-azidophenyl)-propylamino]-benzoic acid and 5-nitro-2-[N-3-(4-azido-2,3,5,6-tetrafluorophenyl)-propylamino]- benzoic acid (FAzNPPB) as photoaffinity labeling agents based on the structure of the widely important Cl- channel blocker 5-nitro-2-(3-phenylpropyl-amino)-benzoic acid (NPPB). The tetrafluoro-substituted aryl azide was found to be a more effective photoinactivating agent than the corresponding protio compound. Using a tritiated version ([3H]FAzNPPB), we demonstrated that photoinactivation of Cl- flux was accompanied by photolabeling of the band 3 protein and membrane lipids. Both processes were diminished in the presence of NPPB and the related arylanthranilate flufenamic acid. Photolabeling resulted in the incorporation of 1.0 +/- 0.2 mol 3H/mol protein in the band 3 integral membrane domain, whereas the cytoplasmic domain was essentially unlabeled. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, photolabeling was found to be the result of partial labeling of at least three different regions of the membrane domain. Based on trypsin proteolysis, reverse-phase high-performance liquid chromatography and electrospray ionization mass spectrometry analysis, it is proposed that one of the sites of photolabeling is the peptide lys-phe-lys (590-592). FAzNPPB is a successful polyfluoro aryl azide photoaffinity labeling agent which may be of further use in studying the diverse effects of arylanthranilates on biological membranes.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D009579 Nitrobenzoates Benzoic acid or benzoic acid esters substituted with one or more nitro groups. Nitrobenzoic Acids,Acids, Nitrobenzoic
D010782 Photolysis Chemical bond cleavage reactions resulting from absorption of radiant energy. Photodegradation
D002712 Chlorides Inorganic compounds derived from hydrochloric acid that contain the Cl- ion. Chloride,Chloride Ion Level,Ion Level, Chloride,Level, Chloride Ion
D004910 Erythrocyte Membrane The semi-permeable outer structure of a red blood cell. It is known as a red cell 'ghost' after HEMOLYSIS. Erythrocyte Ghost,Red Cell Cytoskeleton,Red Cell Ghost,Erythrocyte Cytoskeleton,Cytoskeleton, Erythrocyte,Cytoskeleton, Red Cell,Erythrocyte Cytoskeletons,Erythrocyte Ghosts,Erythrocyte Membranes,Ghost, Erythrocyte,Ghost, Red Cell,Membrane, Erythrocyte,Red Cell Cytoskeletons,Red Cell Ghosts
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000345 Affinity Labels Analogs of those substrates or compounds which bind naturally at the active sites of proteins, enzymes, antibodies, steroids, or physiological receptors. These analogs form a stable covalent bond at the binding site, thereby acting as inhibitors of the proteins or steroids. Affinity Labeling Reagents,Labeling Reagents, Affinity,Labels, Affinity,Reagents, Affinity Labeling
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D001386 Azides Organic or inorganic compounds that contain the -N3 group. Azide
D001457 Anion Exchange Protein 1, Erythrocyte A major integral transmembrane protein of the ERYTHROCYTE MEMBRANE. It is the anion exchanger responsible for electroneutral transporting in CHLORIDE IONS in exchange of BICARBONATE IONS allowing CO2 uptake and transport from tissues to lungs by the red blood cells. Genetic mutations that result in a loss of the protein function have been associated with type 4 HEREDITARY SPHEROCYTOSIS. Anion Transport Protein, Erythrocyte,Band 3 Protein,Erythrocyte Anion Transport Protein,Erythrocyte Membrane Band 3 Protein,AE1 Anion Exchanger,AE1 Chloride-Bicarbonate Exchanger,AE1 Cl- HCO3- Exchanger,AE1 Gene Product,Anion Exchanger 1,Antigens, CD233,Band 3 Anion Transport Protein,Band III Protein,CD233 Antigen,CD233 Antigens,Capnophorin,EPB3 Protein,Erythrocyte Anion Exchanger,Erythrocyte Membrane Anion Transport Protein,Erythrocyte Membrane Protein Band 3, Diego Blood Group,Protein Band 3,SLC4A1 Protein,Solute Carrier Family 4 Member 1,Solute Carrier Family 4, Anion Exchanger, Member 1,AE1 Chloride Bicarbonate Exchanger,AE1 Cl HCO3 Exchanger,Anion Exchanger, Erythrocyte,Antigen, CD233,Chloride-Bicarbonate Exchanger, AE1,Exchanger 1, Anion,Protein, EPB3

Related Publications

B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
June 1992, The Journal of membrane biology,
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
November 1991, Biochimica et biophysica acta,
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
January 2009, Pharmacology, biochemistry, and behavior,
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
January 1996, British journal of pharmacology,
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
December 2009, The Journal of biological chemistry,
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
December 1988, Biochimica et biophysica acta,
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
January 2008, Pharmacology,
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
June 2011, Anatomical record (Hoboken, N.J. : 2007),
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
December 2014, Plant physiology,
B R Branchini, and M H Murtiashaw, and E A Eckman, and L A Egan, and C V Alfano, and J G Stroh
January 2001, Journal of the Society for Gynecologic Investigation,
Copied contents to your clipboard!