Expression, crystallization and preliminary X-ray crystallographic analysis of human agmatinase.
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%.
Kim
Kyoung Hoon
KH
Department of Chemistry, College of Natural Sciences, Seoul National University, Seoul 151-742, South Korea.
Ahn
Hyung Jun
HJ
Kim
Do Jin
DJ
Lee
Hyung Ho
HH
Ha
Jun-Yong
JY
Kim
Hye-Kyung
HK
Yoon
Hye-Jin
HJ
Suh
Se Won
SW
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
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