Preliminary X-ray crystallographic analysis of the catalytic domain of prophenoloxidase activating factor-I.
Clip-domain serine proteases (SPs) have been identified in invertebrates as crucial enzymes that are involved in diverse extracellular signalling pathways. Prophenoloxidase (proPO) activating factor-I (PPAF-I), a catalytically active clip-domain SP, cleaves proPO. To date, no crystal structures of a catalytically active clip-domain SP have been determined. Here, the results of crystallization and preliminary X-ray analysis of the SP domain of PPAF-I are reported. The crystal of the PPAF-I SP domain was obtained using the hanging-drop vapour-diffusion method in a precipitant solution containing 0.15 M lithium sulfate, 30% polyethylene glycol 4000 and 0.1 M Tris-HCl pH 8.0. The crystal diffracts X-rays to 1.7 angstroms resolution using a synchrotron-radiation source. The crystal belongs to space group P2(1)2(1)2(1), with one molecule in the asymmetric unit and unit-cell parameters a = 38.3, b = 53.3, c = 116.6 angstroms, alpha = beta = gamma = 90 degrees. A molecular-replacement solution has been found using kallikrein as a starting model, resulting in an interpretable electron-density map.
Piao
Shunfu
S
College of Pharmacy and Research Institute for Drug Development, Pusan National University, Jangjeon-dong, Geumjeong-gu, Busan 609-735, South Korea.
Jung
Ji Young
JY
Park
Ji Won
JW
Lee
Jaewon
J
Lee
Bok Leul
BL
Ha
Nam Chul
NC
D000595:Amino Acid Sequence; D020134:Catalytic Domain; D004156:Catechol Oxidase / Q000378:metabolism*; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D004792:Enzyme Precursors / Q000378:metabolism*; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000737:chemistry*; D014961:X-Ray Diffraction
D004792:Enzyme Precursors; C038451:pro-phenoloxidase; D004156:Catechol Oxidase; D012697:Serine Endopeptidases; C022435:prephenoloxidase-activating enzyme