title: Expression, crystallization and preliminary X-ray crystallographic analysis of human agmatinase.
abstract: Agmatine, which results from the decarboxylation of L-arginine by arginine decarboxylase, is a metabolic intermediate in the biosynthesis of putresine and higher polyamines (spermidine and spermine). Recent studies indicate that agmatine can have several important biochemical effects in humans, ranging from effects on the central nervous system to cell proliferation in cancer and viral replication. Agmatinase catalyses the hydrolysis of agmatine to putresine and urea and is a major target for drug action and development. The human agmatinase gene encodes a 352-residue protein with a putative mitochondrial targeting sequence at the N-terminus. Human agmatinase (residues Ala36-Val352) has been overexpressed as a fusion with both N- and C-terminal purification tags in Escherichia coli and crystallized in the presence of Mn2+ and 1,6-diaminohexane at 297 K using polyethylene glycol 4000 as a precipitant. X-ray diffraction data were collected at 100 K to 2.49 A from a flash-frozen crystal. The crystals are tetragonal, belonging to space group P4(2), with unit-cell parameters a = b = 114.54, c = 125.65 A, alpha = beta = gamma = 90 degrees. Three monomers are likely to be present in the asymmetric unit, giving a crystal volume per protein weight (VM) of 3.66 A3 Da(-1) and a solvent content of 66.4%.
authors.affiliation: Department of Chemistry, College of Natural Sciences, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Kyoung Hoon
authors.initials: KH
authors.lastname: Kim
authors.forename: Hyung Jun
authors.initials: HJ
authors.lastname: Ahn
authors.forename: Do Jin
authors.initials: DJ
authors.lastname: Kim
authors.forename: Hyung Ho
authors.initials: HH
authors.lastname: Lee
authors.forename: Jun-Yong
authors.initials: JY
authors.lastname: Ha
authors.forename: Hye-Kyung
authors.initials: HK
authors.lastname: Kim
authors.forename: Hye-Jin
authors.initials: HJ
authors.lastname: Yoon
authors.forename: Se Won
authors.initials: SW
authors.lastname: Suh
mesh_terms: D000376:Agmatine / Q000737:chemistry; D001120:Arginine / Q000737:chemistry; D002262:Carboxy-Lyases / Q000737:chemistry; D002384:Catalysis; D018360:Crystallography, X-Ray / Q000379:methods*; D018076:DNA, Complementary / Q000378:metabolism; D003959:Diamines / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007668:Kidney / Q000378:metabolism; D008274:Magnesium / Q000737:chemistry; D008928:Mitochondria / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013095:Spermidine / Q000737:chemistry; D013096:Spermine / Q000737:chemistry; D013696:Temperature; D014508:Urea / Q000737:chemistry; D014512:Ureohydrolases / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: D018076:DNA, Complementary; D003959:Diamines; D011993:Recombinant Fusion Proteins; D013096:Spermine; D011092:Polyethylene Glycols; D000376:Agmatine; D014508:Urea; D001120:Arginine; D014512:Ureohydrolases; C019625:agmatinase; D002262:Carboxy-Lyases; C019722:arginine decarboxylase; D008274:Magnesium; D013095:Spermidine; C007297:1,6-diaminohexane