title: Crystallization of nepenthesin I using a low-pH crystallization screen.
abstract: Nepenthesins are aspartic proteases secreted by carnivorous pitcher plants of the genus Nepenthes. They significantly differ in sequence from other plant aspartic proteases. This difference, which provides more cysteine residues in the structure of nepenthesins, may contribute to their unique stability profile. Recombinantly produced nepenthesin 1 (rNep1) from N. gracilis in complex with pepstatin A was crystallized under two different crystallization conditions using a newly formulated low-pH crystallization screen. The diffraction data were processed to 2.9 and 2.8 Å resolution, respectively. The crystals belonged to space group P212121, with unit-cell parameters a = 86.63, b = 95.90, c = 105.40 Å, α = β = γ = 90° and a = 86.28, b = 97.22, c = 103.78 Å, α = β = γ = 90°, respectively. Matthews coefficient and solvent-content calculations suggest the presence of two molecules of rNep1 in the asymmetric unit. Here, the details of the crystallization experiment and analysis of the X-ray data are reported.
authors.affiliation: Institute of Macromolecular Chemistry CAS, v.v.i., Heyrovského nám. 2/1888, 162 06 Praha 6, Czech Republic.
authors.forename: Karla
authors.initials: K
authors.lastname: Fejfarová
authors.affiliation: Faculty of Science, Charles University in Prague, Albertov 6, 128 44 Praha 2, Czech Republic.
authors.forename: Alan
authors.initials: A
authors.lastname: Kádek
authors.affiliation: Institute of Microbiology CAS, v.v.i., Vídeňská 1083, 142 20 Praha 4, Czech Republic.
authors.forename: Hynek
authors.initials: H
authors.lastname: Mrázek
authors.affiliation: Faculty of Science, Charles University in Prague, Albertov 6, 128 44 Praha 2, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Hausner
authors.affiliation: Faculty of Science, Charles University in Prague, Albertov 6, 128 44 Praha 2, Czech Republic.
authors.forename: Vyacheslav
authors.initials: V
authors.lastname: Tretyachenko
authors.affiliation: Institute of Macromolecular Chemistry CAS, v.v.i., Heyrovského nám. 2/1888, 162 06 Praha 6, Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Koval'
authors.affiliation: Faculty of Science, Charles University in Prague, Albertov 6, 128 44 Praha 2, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Man
authors.affiliation: Institute of Biotechnology CAS, v.v.i., Vídeňská 1083, 142 20 Praha 4, Czech Republic.
authors.forename: Jindřich
authors.initials: J
authors.lastname: Hašek
authors.affiliation: Institute of Macromolecular Chemistry CAS, v.v.i., Heyrovského nám. 2/1888, 162 06 Praha 6, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Dohnálek
mesh_terms: D016282:Aspartic Acid Endopeptidases / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D006863:Hydrogen-Ion Concentration; D019684:Magnoliopsida / Q000201:enzymology; D010436:Pepstatins / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry*
chemical_list: D010436:Pepstatins; D010940:Plant Proteins; D016282:Aspartic Acid Endopeptidases; C031375:pepstatin
keywords: Nepenthes gracilis; aspartic proteases; low-pH crystallization screen; nepenthesins