Crystallization of nepenthesin I using a low-pH crystallization screen

• Published in: Acta crystallographica. Section F, Structural biology communications Vol. 72; no. 1; pp. 24 - 28
• Main Authors: Fejfarová, Karla, Kádek, Alan, Mrázek, Hynek, Hausner, Jiří, Tretyachenko, Vyacheslav, Koval', Tomáš, Man, Petr, Hašek, Jindřich, Dohnálek, Jan
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.01.2016; Wiley Subscription Services, Inc
• Subjects: Aspartic Acid Endopeptidases - chemistry; aspartic proteases; Asymmetry; Biology; Crystallization; Crystallography, X-Ray; Crystals; Diffraction; Hydrogen-Ion Concentration; low-pH crystallization screen; Magnoliopsida - enzymology; Mathematical analysis; Nepenthes gracilis; nepenthesins; Pepstatins - chemistry; Plant Proteins - chemistry; Plants (organisms); Protease; Research Communications; Screens; nepenthesins; low-pH crystallization screen; Nepenthes gracilis; aspartic proteases
• ISSN: 2053-230X; 2053-230X
• DOI: 10.1107/S2053230X15022323

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.