title: Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human histidine triad nucleotide-binding protein 2 (hHINT2).
abstract: Histidine triad nucleotide-binding protein 2 (HINT2) is a mitochondrial adenosine phosphoramidase mainly expressed in the pancreas, liver and adrenal gland. HINT2 possibly plays a role in apoptosis, as well as being involved in steroid biosynthesis, hepatic lipid metabolism and regulation of hepatic mitochondria function. The expression level of HINT2 is significantly down-regulated in hepatocellular carcinoma patients. To date, endogenous substrates for this enzyme, as well as the three-dimensional structure of human HINT2, are unknown. In this study, human HINT2 was cloned, overexpressed in Escherichia coli and purified. Crystallization was performed at 278 K using PEG 4000 as the main precipitant; the crystals, which belonged to the tetragonal space group P41212 with unit-cell parameters a = b = 76.38, c = 133.25 Å, diffracted to 2.83 Å resolution. Assuming two molecules in the asymmetric unit, the Matthews coefficient and the solvent content were calculated to be 2.63 Å(3) Da(-1) and 53.27%, respectively.
authors.affiliation: Department of Bioorganic Chemistry, Centre of Molecular and Macromolecular Studies, Sienkiewicza 112, 90-363 Łódź, Poland. rdolot@cbmm.lodz.pl
authors.forename: Rafał
authors.initials: R
authors.lastname: Dolot
authors.forename: Artur
authors.initials: A
authors.lastname: Włodarczyk
authors.forename: Grzegorz D
authors.initials: GD
authors.lastname: Bujacz
authors.forename: Barbara
authors.initials: B
authors.lastname: Nawrot
mesh_terms: D000595:Amino Acid Sequence; D002478:Cells, Cultured; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012997:Solvents / Q000737:chemistry
chemical_list: C533603:Hint2 protein, human; D024101:Mitochondrial Proteins; D011994:Recombinant Proteins; D012997:Solvents
keywords: HINT; histidine triad nucleotide-binding protein