Expression, crystallization and preliminary crystallographic data analysis of PigI, a putative L-prolyl-AMP ligase from the prodigiosin synthetic pathway in Serratia

Acta Crystallogr F Struct Biol Commun. 2014 May;70(Pt 5):624-7. doi: 10.1107/S2053230X14005780. Epub 2014 Apr 17.

Abstract

Prodigiosin, a member of the prodiginines, is a tripyrrole red pigment synthesized by Serratia and some other microbes. A bifurcated biosynthesis pathway of prodigiosin has been proposed in Serratia in which MBC (4-methoxy-2,2'-bipyrrole-5-carbaldehyde) and MAP (2-methyl-3-N-amyl-pyrrole) are synthesized separately and then condensed by PigC to form prodigiosin. The first step for the synthesis of MBC is the activation of L-proline by PigI, but its catalytic mechanism has remained elusive. To elucidate its mechanism, recombinant PigI was purified and crystallized. Crystals obtained by the sitting-drop method belonged to space group P1 and diffracted to 2.0 Å resolution, with unit-cell parameters a = 51.2, b = 62.8, c = 91.3 Å, α = 105.1, β = 90.1, γ = 92.2°. Matthews coefficient analysis suggested two molecules in the asymmetric unit, with a VM of 2.6 Å(3) Da(-1) and a solvent content of 52.69%.

Keywords: PigI; Serratia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization
  • Crystallography, X-Ray
  • DNA Ligases / biosynthesis*
  • DNA Ligases / chemistry*
  • Gene Expression Regulation, Enzymologic*
  • Prodigiosin / biosynthesis*
  • Prodigiosin / chemistry*
  • Serratia / enzymology*
  • Signal Transduction / physiology

Substances

  • Ampligase
  • DNA Ligases
  • Prodigiosin