Crystal structure of the yeast nicotinamidase Pnc1p

Gang Hu, Alexander B. Taylor, Lee McAlister-Henn, P. John Hart

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The yeast nicotinamidase Pnc1p acts in transcriptional silencing by reducing levels of nicotinamide, an inhibitor of the histone deacetylase Sir2p. The Pnc1p structure was determined at 2.9 Å resolution using MAD and MIRAS phasing methods after inadvertent crystallization during the pursuit of the structure of histidine-tagged yeast isocitrate dehydrogenase (IDH). Pnc1p displays a cluster of surface histidine residues likely responsible for its co-fractionation with IDH from Ni2+-coupled chromatography resins. Researchers expressing histidine-tagged proteins in yeast should be aware of the propensity of Pnc1p to crystallize, even when overwhelmed in concentration by the protein of interest. The protein assembles into extended helical arrays interwoven to form an unusually robust, yet porous superstructure. Comparison of the Pnc1p structure with those of three homologous bacterial proteins reveals a common core fold punctuated by amino acid insertions unique to each protein. These insertions mediate the self-interactions that define the distinct higher order oligomeric states attained by these molecules. Pnc1p also acts on pyrazinamide, a substrate analog converted by the nicotinamidase from Mycobacterium tuberculosis into a product toxic to that organism. However, we find no evidence for detrimental effects of the drug on yeast cell growth.

Original languageEnglish (US)
Pages (from-to)66-75
Number of pages10
JournalArchives of Biochemistry and Biophysics
Volume461
Issue number1
DOIs
StatePublished - May 1 2007

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Nicotinamidase
Histidine
Yeast
Isocitrate Dehydrogenase
Yeasts
Crystal structure
Pyrazinamide
Proteins
Histone Deacetylase Inhibitors
Fungal Proteins
Bacterial Proteins
Niacinamide
Poisons
Crystallization
Mycobacterium tuberculosis
Chromatography
Cell growth
Fractionation
Research Personnel
Amino Acids

Keywords

  • Kinetic analyses
  • Multiple isomorphous replacement
  • Multiwavelength anomalous diffraction
  • NAD
  • Nicotinamidase
  • Sir2p
  • X-ray crystallography

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Crystal structure of the yeast nicotinamidase Pnc1p. / Hu, Gang; Taylor, Alexander B.; McAlister-Henn, Lee; Hart, P. John.

In: Archives of Biochemistry and Biophysics, Vol. 461, No. 1, 01.05.2007, p. 66-75.

Research output: Contribution to journalArticle

Hu, Gang ; Taylor, Alexander B. ; McAlister-Henn, Lee ; Hart, P. John. / Crystal structure of the yeast nicotinamidase Pnc1p. In: Archives of Biochemistry and Biophysics. 2007 ; Vol. 461, No. 1. pp. 66-75.
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