Aspartate aminotransferase: An old dog teaches new tricks

Michael D. Toney

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Aspartate aminotransferase (AAT) is a prototypical pyridoxal 5′-phosphate (PLP) dependent enzyme that catalyzes the reversible interconversion of l-aspartate and α-ketoglutarate with oxalacetate and l-glutamate via a ping-pong catalytic cycle in which the pyridoxamine 5′-phosphate enzyme form is an intermediate. There is a bountiful literature on AAT that spans approximately 60 years, and much fundamental mechanistic information on PLP dependent reactions has been gained from its study. Here, we review our recent work on AAT, where we again used it as a test bed for fundamental concepts in PLP chemistry. First, we discuss the role that coenzyme protonation state plays in controlling reaction specificity, then ground state destabilization via hyperconjugation in the external aldimine intermediate is examined. The third topic is light enhancement of catalysis of Cα-H deprotonation by PLP in solution and in AAT, which occurs through a triplet state of the external aldimine intermediate. Lastly, we consider recent advances in our analyses of enzyme multiple sequence alignments for the purpose of predicting mutations that are required to interconvert structurally similar but catalytically distinct enzymes, and the application of our program JANUS to the conversion of AAT into tyrosine aminotransferase.

Original languageEnglish (US)
Pages (from-to)119-127
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume544
DOIs
StatePublished - Feb 15 2014

Fingerprint

Aspartate Aminotransferases
Pyridoxal Phosphate
Dogs
Enzymes
Pyridoxamine
Tyrosine Transaminase
Deprotonation
Sequence Alignment
Protonation
Coenzymes
Catalysis
Aspartic Acid
Ground state
Glutamic Acid
Phosphates
Light
Mutation

Keywords

  • Asparate aminotransferase
  • Binding isotope effect
  • Bioinformatics
  • Coenzyme analogue
  • Light activation
  • Nitrogen metabolism
  • Pyridoxal phosphate
  • Vitamin B6

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Aspartate aminotransferase : An old dog teaches new tricks. / Toney, Michael D.

In: Archives of Biochemistry and Biophysics, Vol. 544, 15.02.2014, p. 119-127.

Research output: Contribution to journalArticle

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