Fine tuning of coenzyme specificity in family 2 aldo-keto reductases revealed by crystal structures of the Lys-274 → Arg mutant of Candida tenuis xylose reductase (AKR2B5) bound to NAD+ and NADP+

Stefan Leitgeb, Barbara Petschacher, David K. Wilson, Bernd Nidetzky

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

Aldo-keto reductases of family 2 employ single site replacement Lys → Arg to switch their cosubstrate preference from NADPH to NADH. X-ray crystal structures of Lys-274 → Arg mutant of Candida tenuis xylose reductase (AKR2B5) bound to NAD+ and NADP+ were determined at a resolution of 2.4 and 2.3 Å, respectively. Due to steric conflicts in the NADP+-bound form, the arginine side chain must rotate away from the position of the original lysine side chain, thereby disrupting a network of direct and water-mediated interactions between Glu-227, Lys-274 and the cofactor 2′-phosphate and 3′-hydroxy groups. Because anchoring contacts of its Glu-227 are lost, the coenzyme-enfolding loop that becomes ordered upon binding of NAD(P)+ in the wild-type remains partly disordered in the NADP+-bound mutant. The results delineate a catalytic reaction profile for the mutant in comparison to wild-type.

Original languageEnglish (US)
Pages (from-to)763-767
Number of pages5
JournalFEBS Letters
Volume579
Issue number3
DOIs
StatePublished - Jan 31 2005

Keywords

  • Dual coenzyme specificity
  • Loop ordering
  • Selectivity
  • Xylose fermentation

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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