Pig gastric (H+ + K+)-ATPase. Lys-497 conserved in cation transporting ATPases is modified with pyridoxal 5'-phosphate

Shigehiko Tamura, M. Tagaya, M. Maeda, M. Futai

Research output: Contribution to journalArticle

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Abstract

Pig gastric (H+ + K+)-ATPase can be covalently modified with pyridoxal 5'-phosphate (PLP) (about 1 mol/mol enzyme), and this modification is not observed in the presence of ATP, suggesting that PLP binds to a specific Lys residue in the ATP binding site or the region in its vicinity (Maeda, M., Tagaya, M., and Futai, M. (1988) J. Biol. Chem. 263, 3652-3656). The peptides labeled with radioactive PLP could be released from the gastric membrane vesicles quantitatively by chymotrypsin treatment, and two peptides were purified by high performance liquid chromatographies. These peptides were not obtained from vesicles incubated with PLP in the presence of ATP. The sequences of the two peptides were NH2-Asn-Ser-Thr-Asn-Lys-Phe-COOH and NH2-Ser-Thr-Asn-Lys-Phe-COOH, exactly corresponding to residues 493-498 and 494-498, respectively, of pig gastric (H+ + K+)-ATPase sequenced recently (Maeda, M., Ishizaki, J., and Futai, M. (1988) Biochem. Biophys. Res. Commun. 157, 203-209). Lys-497 was concluded to be the binding site of PLP, as pyridoxyl-Lys was identified at the corresponding position. This Lys residue is conserved in (Na+ + K+)- and Ca2+-ATPases. The possible amino acid residues in the catalytic site of gastric (H+ + K+)-ATPase are discussed.

Original languageEnglish
Pages (from-to)8580-8584
Number of pages5
JournalJournal of Biological Chemistry
Volume264
Issue number15
Publication statusPublished - 1989
Externally publishedYes

Fingerprint

H(+)-K(+)-Exchanging ATPase
Pyridoxal Phosphate
Adenosine Triphosphatases
Cations
Swine
Peptides
Adenosine Triphosphate
Binding Sites
Calcium-Transporting ATPases
Chymotrypsin
High performance liquid chromatography
Catalytic Domain
Stomach
High Pressure Liquid Chromatography
Membranes
Amino Acids
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Pig gastric (H+ + K+)-ATPase. Lys-497 conserved in cation transporting ATPases is modified with pyridoxal 5'-phosphate. / Tamura, Shigehiko; Tagaya, M.; Maeda, M.; Futai, M.

In: Journal of Biological Chemistry, Vol. 264, No. 15, 1989, p. 8580-8584.

Research output: Contribution to journalArticle

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T1 - Pig gastric (H+ + K+)-ATPase. Lys-497 conserved in cation transporting ATPases is modified with pyridoxal 5'-phosphate

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AU - Futai, M.

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