Crystallization of the effector-binding domains of BenM and CatM, LysR-type transcriptional regulators from Acinetobacter sp. ADP1

Acta Crystallogr D Biol Crystallogr. 2004 Jan;60(Pt 1):105-8. doi: 10.1107/s0907444903021589. Epub 2003 Dec 18.

Abstract

BenM, a member of the LysR-type family of transcriptional regulators, controls genes for benzoate degradation in the Gram-negative bacterium Acinetobacter sp. strain ADP1. Recent studies show that BenM activates benABCDE expression synergistically in response to two effector ligands: cis,cis-muconate (CCM) and benzoate. As an initial step in investigating the structural basis of dual effector response, the effector-binding domain of BenM (BenM-EBD) was crystallized by the microbatch-under-oil technique with conditions optimized from high-throughput screens performed by the Hauptman-Woodward Institute. Data-collection quality crystals of BenM-EBD belonged to space group P2(1)2(1)2(1), diffracted to 2.3 A and had unit-cell parameters a = 65.64, b = 66.34, c = 117.46 A. The influence of effector ligands on crystal formation was also evaluated. The presence of benzoate or CCM impaired the formation of crystals. The presence of both effectors together resulted in a dramatic decrease in the production of crystals. The effector-binding domain of CatM, a homolog of BenM, was also crystallized.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acinetobacter / chemistry*
  • Acinetobacter / metabolism
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism
  • Benzoates / chemistry
  • Benzoates / metabolism
  • Crystallization
  • Crystallography, X-Ray
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / metabolism
  • Sorbic Acid / analogs & derivatives*
  • Sorbic Acid / chemistry
  • Sorbic Acid / metabolism
  • Transcription Factors / chemistry*
  • Transcription Factors / metabolism

Substances

  • Bacterial Proteins
  • BenM protein, Acinetobacter
  • Benzoates
  • DNA-Binding Proteins
  • Transcription Factors
  • catM protein, Acinetobacter
  • muconic acid
  • Sorbic Acid