Crystallization of nepenthesin I using a low-pH crystallization screen

0457610 - ÚMCH 2017 RIV GB eng J - Journal Article
Fejfarová, Karla - Kádek, Alan - Mrázek, Hynek - Hausner, Jiří - Tretyachenko, V. - Koval, Tomáš - Man, Petr - Hašek, J. - Dohnálek, Jan
Crystallization of nepenthesin I using a low-pH crystallization screen.
Acta Crystallographica Section F-Structural Biology Communications. Roč. 72, č. 1 (2016), s. 24-28. E-ISSN 2053-230X
R&D Projects: GA MŠMT(CZ) EE2.3.30.0029; GA MŠMT(CZ) ED1.1.00/02.0109; GA ČR GAP206/12/0503
Institutional support: RVO:61389013 ; RVO:61388971
Keywords : aspartic proteases * nepenthesins * Nepenthes gracilis
Subject RIV: CE - Biochemistry; EB - Genetics ; Molecular Biology (MBU-M)
Impact factor: 0.799, year: 2016

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 A resolution, respectively. The crystals belonged to space group P212121, with unit-cell parameters a = 86.63, b = 95.90, c = 105.40 A, [alpha] = [beta] = [gamma] = 90° and a = 86.28, b = 97.22, c = 103.78 A, [alpha] = [beta] = [gamma] = 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.
Permanent Link: http://hdl.handle.net/11104/0258010