Regulation of phenylalanine hydroxylase: Conformational changes upon phenylalanine binding detected by hydrogen/deuterium exchange and mass spectrometry

Jun Li, Lawrence J. Dangott, Paul F. Fitzpatrick

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

Abstract

Phenylalanine acts as an allosteric activator of the tetrahydropterin- dependent enzyme phenylalanine hydroxylase. Hydrogen/deuterium exchange monitored by mass spectrometry has been used to gain insight into local conformational changes accompanying activation of rat phenylalanine hydroxylase by phenylalanine. Peptides in the regulatory and catalytic domains that lie in the interface between these two domains show large increases in the extent of deuterium incorporation from solvent in the presence of phenylalanine. In contrast, the effects of phenylalanine on the exchange kinetics of a mutant enzyme lacking the regulatory domain are limited to peptides surrounding the binding site for the amino acid substrate. These results support a model in which the N-terminus of the protein acts as an inhibitory peptide, with phenylalanine binding causing a conformational change in the regulatory domain that alters the interaction between the catalytic and regulatory domains.

Original languageEnglish (US)
Pages (from-to)3327-3335
Number of pages9
JournalBiochemistry
Volume49
Issue number15
DOIs
StatePublished - Apr 20 2010

ASJC Scopus subject areas

  • Biochemistry

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