Role for an essential tyrosine in peptide amidation

Mithu De, Joseph Bell, Ninian Blackburn, Richard E. Mains, Betty A. Eipper

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The catalytic core of the peptidyl-α-hydroxyglycine α-amidating lyase (PAL) domain of peptidylglycine α-amidating monooxygenase was investigated with respect to its ability to function as a ureidoglycolate lyase and the identity and role of its bound metal ions. The purified PAL catalytic core (PALcc) contains molar equivalents of calcium and zinc along with substoichiometric amounts of iron and functions as a ureidoglycolate lyase. Limiting iron availability in the cells synthesizing PALcc reduces the specific activity of the enzyme produced. Concentrated samples of native PALcc have an absorption maximum at 560 nm, suggestive of a phenolate-Fe(III) charge transfer complex. An essential role for a Tyr residue was confirmed by elimination of PAL activity following site-directed mutagenesis. Purified PALcc in which the only conserved Tyr residue (Tyr654) was mutated to Phe was secreted normally, but was catalytically inactive and lacked bound iron and bound zinc. Our data demonstrate an essential role for Tyr654 and suggest that it serves as an Fe(III) ligand in an essential iron-zinc bimetallic site.

Original languageEnglish (US)
Pages (from-to)20873-20882
Number of pages10
JournalJournal of Biological Chemistry
Volume281
Issue number30
DOIs
StatePublished - Jul 28 2006

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Lyases
Tyrosine
Catalytic Domain
Iron
Peptides
Zinc
Mutagenesis
Mixed Function Oxygenases
Site-Directed Mutagenesis
Metal ions
Charge transfer
Metals
Availability
Ions
Ligands
Calcium
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Role for an essential tyrosine in peptide amidation. / De, Mithu; Bell, Joseph; Blackburn, Ninian; Mains, Richard E.; Eipper, Betty A.

In: Journal of Biological Chemistry, Vol. 281, No. 30, 28.07.2006, p. 20873-20882.

Research output: Contribution to journalArticle

De, Mithu ; Bell, Joseph ; Blackburn, Ninian ; Mains, Richard E. ; Eipper, Betty A. / Role for an essential tyrosine in peptide amidation. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 30. pp. 20873-20882.
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