title: Crystallization and preliminary X-ray diffraction analysis of Xaa-Pro dipeptidase from Xanthomonas campestris.
abstract: Xaa-Pro dipeptidase (XPD; prolidase; EC 3.4.13.9) specifically hydrolyzes dipeptides with a prolyl residue at the carboxy-terminus. Xanthomonas spp. possess two different isoforms of XPD (48 and 43 kDa) which share ∼24% sequence identity. The XPD of 43 kDa in size (XPD43) from Xanthomonas spp. is unusual as it lacks the strictly conserved tyrosine residue (equivalent to Tyr387 in Escherichia coli aminopeptidase P) that is suggested to be important in the proton-shuttle transfer required for catalysis in the M24B (MEROPS) family. Here, the crystallization and preliminary X-ray analysis of XPD43 from X. campestris (GenBank accession No. NP_637763) are reported. Recombinant XPD43 was crystallized using the microbatch-under-oil technique. Diffraction data were collected on the recently commissioned protein crystallography beamline (PX-BL21) at the Indian synchrotron (Indus-2, 2.5 GeV) to 1.83 Å resolution with 100% completeness. The crystal belonged to space group P212121, with unit-cell parameters a = 84.32, b = 105.51, c = 111.35 Å. Two monomers are expected to be present in the asymmetric unit of the crystal, corresponding to a solvent content of 58%. Structural analysis of XPD43 will provide new insights into the role of the conserved residues in catalysis in the M24B family.
authors.affiliation: High Pressure and Synchrotron Radiation Physics Division, Bhabha Atomic Research Centre, Trombay, Mumbai, Maharashtra 400 085, India.
authors.forename: Ashwani
authors.initials: A
authors.lastname: Kumar
authors.affiliation: High Pressure and Synchrotron Radiation Physics Division, Bhabha Atomic Research Centre, Trombay, Mumbai, Maharashtra 400 085, India.
authors.forename: Venkata Narayana
authors.initials: VN
authors.lastname: Are
authors.affiliation: High Pressure and Synchrotron Radiation Physics Division, Bhabha Atomic Research Centre, Trombay, Mumbai, Maharashtra 400 085, India.
authors.forename: Biplab
authors.initials: B
authors.lastname: Ghosh
authors.affiliation: High Pressure and Synchrotron Radiation Physics Division, Bhabha Atomic Research Centre, Trombay, Mumbai, Maharashtra 400 085, India.
authors.forename: Utsavi
authors.initials: U
authors.lastname: Agrawal
authors.affiliation: Food Technology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, Maharashtra 400 085, India.
authors.forename: Sahayog N
authors.initials: SN
authors.lastname: Jamdar
authors.affiliation: High Pressure and Synchrotron Radiation Physics Division, Bhabha Atomic Research Centre, Trombay, Mumbai, Maharashtra 400 085, India.
authors.forename: Ravindra D
authors.initials: RD
authors.lastname: Makde
authors.affiliation: High Pressure and Synchrotron Radiation Physics Division, Bhabha Atomic Research Centre, Trombay, Mumbai, Maharashtra 400 085, India.
authors.forename: Surinder M
authors.initials: SM
authors.lastname: Sharma
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D017931:DNA Primers; D004150:Dipeptidases / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D016959:Xanthomonas campestris / Q000201:enzymology*
chemical_list: D017931:DNA Primers; D004150:Dipeptidases; C038937:dipeptidase
keywords: Xaa-Pro dipeptidase; Xanthomonas campestris; prolidase