Crystallization of Pseudomonas aeruginosa AHL synthase LasI using beta-turn crystal engineering.
In Gram-negative bacteria, intercellular communication and virulence regulation is mediated by the diffusible chemical signal acyl-homoserine-L-lactone (AHL). The AHL synthase enzymes produce a variety of AHLs from the substrates S-adenosyl-L-methionine and acyl-acyl carrier protein. LasI, the AHL synthase from Pseudomonas aeruginosa, has low solubility and has failed to crystallize despite extensive crystallization trials. Based on the previously determined structure of the AHL synthase EsaI, active soluble LasI was produced by re-engineering residues in a tight turn to produce a type I' beta-turn. The resulting protein is active, more stable than the wild-type LasI and has been crystallized in the cubic space group F23, with unit-cell parameters a = b = c = 154.90 A.
Gould
Ty A
TA
Department of Pharmacology, The University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, CO 80262, USA.
Watson
William T
WT
Choi
Kyoung Hee
KH
Schweizer
Herbert P
HP
Churchill
Mair E A
ME
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D008025:Ligases / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017433:Protein Structure, Secondary; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D016415:Sequence Alignment
D001426:Bacterial Proteins; C088386:homoserine lactone; D008025:Ligases; C449816:N-acylhomoserine lactone synthase; D015107:4-Butyrolactone