title: Expression, purification, crystallization and preliminary X-ray diffraction analysis of acylpeptide hydrolase from Deinococcus radiodurans.
abstract: Acylpeptide hydrolase (APH; EC 3.4.19.1), which belongs to the S9 family of serine peptidases (MEROPS), catalyzes the removal of an N-acylated amino acid from a blocked peptide. The role of this enzyme in mammalian cells has been suggested to be in the clearance of oxidatively damaged proteins as well as in the degradation of the β-amyloid peptides implicated in Alzheimer's disease. Detailed structural information for the enzyme has been reported from two thermophilic archaea; both of the archaeal APHs share a similar monomeric structure. However, the mechanisms of substrate selectivity and active-site accessibility are totally different and are determined by inter-domain flexibility or the oligomeric structure. An APH homologue from a bacterium, Deinococcus radiodurans (APHdr), has been crystallized using microbatch-under-oil employing the random microseed matrix screening method. The protein crystallized in space group P21, with unit-cell parameters a = 77.6, b = 189.6, c = 120.4 Å, β = 108.4°. A Matthews coefficient of 2.89 Å(3) Da(-1) corresponds to four monomers, each with a molecular mass of ∼73 kDa, in the asymmetric unit. The APHdr structure will reveal the mechanisms of substrate selectivity and active-site accessibility in the bacterial enzyme. It will also be helpful in elucidating the functional role of this enzyme in D. radiodurans.
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: 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: Pooja
authors.initials: P
authors.lastname: Yadav
authors.affiliation: School of Biochemistry, Devi Ahilya University, Indore 452 001, India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Bhatnagar
authors.affiliation: Food Technology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, Maharashtra 400085, 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
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D017931:DNA Primers; D034301:Deinococcus / Q000201:enzymology*; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011487:Protein Conformation
chemical_list: D017931:DNA Primers; D010447:Peptide Hydrolases; C038220:acylaminoacyl-peptidase
keywords: Deinococcus radiodurans; S9 serine peptidase; acylpeptide hydrolase