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Crystallization of nepenthesin I using a low‐pH crystallization screen
Author(s) -
Fejfarová Karla,
Kádek Alan,
Mrázek Hynek,
Hausner Jiří,
Tretyachenko Vyacheslav,
Koval' Tomáš,
Man Petr,
Hašek Jindřich,
Dohnálek Jan
Publication year - 2016
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x15022323
Subject(s) - crystallization , pepstatin , proteases , crystallography , molecule , solvent , chemistry , stereochemistry , biochemistry , organic chemistry , enzyme , protease
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 P 2 1 2 1 2 1 , 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.