Crystallization of a newly discovered histidine acid phosphatase from Francisella tularensis.
Francisella tularensis is a highly infectious bacterial pathogen that is considered by the Centers for Disease Control and Prevention to be a potential bioterrorism weapon. Here, the crystallization of a 37.2 kDa phosphatase encoded by the genome of F. tularensis subsp. holarctica live vaccine strain is reported. This enzyme shares 41% amino-acid sequence identity with Legionella pneumophila major acid phosphatase and contains the RHGXRXP motif that is characteristic of the histidine acid phosphatase family. Large diffraction-quality crystals were grown in the presence of Tacsimate, HEPES and PEG 3350. The crystals belong to space group P4(1)2(1)2, with unit-cell parameters a = 61.96, c = 210.78 A. The asymmetric unit is predicted to contain one protein molecule, with a solvent content of 53%. A 1.75 A resolution native data set was recorded at beamline 4.2.2 of the Lawrence Berkeley National Laboratory Advanced Light Source. Molecular-replacement trials using the human prostatic acid phosphatase structure as the search model (28% amino-acid sequence identity) did not produce a satisfactory solution. Therefore, the structure of F. tularensis histidine acid phosphatase will be determined by multiwavelength anomalous dispersion phasing using a selenomethionyl derivative.
Felts
Richard L
RL
Department of Chemistry, University of Missouri-Columbia, Columbia, Missouri 65211, USA.
Reilly
Thomas J
TJ
Calcutt
Michael J
MJ
Tanner
John J
JJ
D000135:Acid Phosphatase / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D005604:Francisella tularensis / Q000201:enzymology*; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D008297:Male; D011467:Prostate / Q000201:enzymology; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry
D001426:Bacterial Proteins; D006639:Histidine; D000135:Acid Phosphatase; C428439:prostatic acid phosphatase; D017027:Protein Tyrosine Phosphatases