title: Localization of the pyridoxal phosphate binding site at the COOH-terminal region of erythrocyte band 3 protein.
abstract: A human erythrocyte Band 3 peptide, affinity labeled with pyridoxal phosphate, was purified by a combination of gel permeation and reverse-phase high performance liquid chromatography. The amino acid sequence of the transmembrane peptide was determined by sequencing subfragments of the peptide obtained from lysyl endopeptidase and staphylococcal proteinase V8 digestions. When a peptide containing the COOH-terminal of human erythrocyte Band 3 was also purified and sequenced, the affinity-labeled peptide was found to be located close to the COOH-terminal of Band 3, where it could be aligned with amino acid residues 852-927 of a murine erythrocyte Band 3, deduced from a nucleotide sequence of a cDNA clone (Kopito, R. R., and Lodish, H. F. (1985) Nature 316, 234-238). The amino acid sequence of the COOH-terminal region was highly homologous to that of murine Band 3. As a result, the sequence of the COOH-terminal peptide of Band 3 was established as follows. (Formula: see text). The pyridoxal phosphate binding site was identified as Lys-18 which corresponded to Lys-869 of the deduced sequence. It appears that the COOH-terminal region of Band 3 constitutes at least a part of the active center for anion transport in human erythrocyte membranes.
authors: Department of Biochemistry, Fukuoka University School of Medicine, Japan.
authors: Y
authors: Y
authors: Kawano
authors: K
authors: K
authors: Okubo
authors: F
authors: F
authors: Tokunaga
authors: T
authors: T
authors: Miyata
authors: S
authors: S
authors: Iwanaga
authors: N
authors: N
authors: Hamasaki
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D011732:Pyridoxal Phosphate / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D001457:Anion Exchange Protein 1, Erythrocyte; D011732:Pyridoxal Phosphate; D003488:Cyanogen Bromide