Expression, purification and crystallization of class C acid phosphatases from Francisella tularensis and Pasteurella multocida.
Class C nonspecific acid phosphatases are bacterial enzymes that are secreted across the cytoplasmic membrane and hydrolyze a variety of phosphomonoesters at acidic pH. These enzymes are of interest for the development of improved vaccines and clinical diagnostic methods. In one case, the category A pathogen Francisella tularensis, the class C phosphatase plays a role in bacterial fitness. Here, the cloning, expression, purification and crystallization methods for the class C acid phosphatases from F. tularensis and Pasteurella multocida are reported. Crystals of the F. tularensis enzyme diffracted to 2.0 A resolution and belonged to space group C222(1), with one enzyme molecule in the asymmetric unit. Crystals of the P. multocida enzyme diffracted to 1.85 A resolution and belonged to space group C2, with three molecules in the asymmetric unit. Diffraction patterns from crystals of the P. multocida enzyme exhibited multiple interpenetrating reciprocal-space lattices, indicating epitaxial twinning. Despite this aberrance, autoindexing was robust and the data could be satisfactorily processed to 1.85 A resolution using MOSFLM and SCALA.
Singh
Harkewal
H
Department of Chemistry, University of Missouri-Columbia, 65211, USA.
Felts
Richard L
RL
Ma
Li
L
Malinski
Thomas J
TJ
Calcutt
Michael J
MJ
Reilly
Thomas J
TJ
Tanner
John J
JJ
D000135:Acid Phosphatase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D003460:Crystallization; D018360:Crystallography, X-Ray; D005604:Francisella tularensis / Q000201:enzymology*; D016979:Pasteurella multocida / Q000201:enzymology*
D000135:Acid Phosphatase