Crystallization of the effector-binding domains of BenM and CatM, LysR-type transcriptional regulators from Acinetobacter sp. ADP1.
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.
Clark
Todd
T
Department of Microbiology, University of Georgia, Athens, GA, USA.
Haddad
Sandra
S
Neidle
Ellen
E
Momany
Cory
C
D000150:Acinetobacter / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001565:Benzoates / Q000737:chemistry / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D013011:Sorbic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism
D001426:Bacterial Proteins; C112178:BenM protein, Acinetobacter; D001565:Benzoates; D004268:DNA-Binding Proteins; D014157:Transcription Factors; C061264:catM protein, Acinetobacter; C005939:muconic acid; D013011:Sorbic Acid