title: Overexpression and preliminary X-ray crystallographic analysis of prophenoloxidase activating factor II, a clip domain family of serine proteases.
abstract: A clip domain family of serine proteases has been identified in invertebrates as a crucial enzyme involved in diverse biological processes including immune responses and embryonic development. Although these proteins contain at least one clip domain at the N-terminal of the serine protease domain, the roles and three-dimensional structure of the clip domain are unknown. Prophenoloxidase activating factor-II (PPAF-II), a clip domain family of serine proteases, derived from the beetle Holotrichia diomphalia larvae, was overexpressed in the baculovirus system, and crystallized using the hanging-drop vapor-diffusion method. High-quality single crystals of PPAF-II were obtained in a precipitant solution containing 0.15 M ammonium sulfate, 1.25 M lithium sulfate monohydrate, and 0.1 M sodium citrate dehydrate (pH 5.5). These crystals belong to space group C2 with unit-cell parameters a=107.84, b=76.78, c=70.49 A and beta=113.93 degrees , and contain one or two molecules in the asymmetric unit. Determination of the three-dimensional structure of PPAF-II would clarify the functions of the clip domains.
authors.affiliation: National Research Laboratory of Defense Proteins, College of Pharmacy, Pusan National University, Jangjeon Dong, Geumjeong Gu, Busan 609-735, Korea.
authors.forename: Shunfu
authors.initials: S
authors.lastname: Piao
authors.forename: Deukmi
authors.initials: D
authors.lastname: Kim
authors.forename: Ji
authors.initials: J
authors.lastname: Won Park
authors.forename: Bok
authors.initials: B
authors.lastname: Leul Lee
authors.forename: Nam-Chul
authors.initials: NC
authors.lastname: Ha
mesh_terms: D018360:Crystallography, X-Ray; D005822:Genetic Vectors; D008958:Models, Molecular; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D012697:Serine Endopeptidases; C022435:prephenoloxidase-activating enzyme