Crystallization of nepenthesin I using a low-pH crystallization screen
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
26750480
PubMed Central
PMC4708046
DOI
10.1107/s2053230x15022323
PII: S2053230X15022323
Knihovny.cz E-resources
- Keywords
- Nepenthes gracilis, aspartic proteases, low-pH crystallization screen, nepenthesins,
- MeSH
- Aspartic Acid Endopeptidases chemistry MeSH
- Hydrogen-Ion Concentration MeSH
- Crystallization MeSH
- Crystallography, X-Ray MeSH
- Magnoliopsida enzymology MeSH
- Pepstatins chemistry MeSH
- Plant Proteins chemistry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Aspartic Acid Endopeptidases MeSH
- pepstatin MeSH Browser
- Pepstatins MeSH
- Plant Proteins MeSH
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.
Faculty of Science Charles University Prague Albertov 6 128 44 Praha 2 Czech Republic
Institute of Biotechnology CAS v v i Vídeňská 1083 142 20 Praha 4 Czech Republic
Institute of Macromolecular Chemistry CAS v v i Heyrovského nám 2 1888 162 06 Praha 6 Czech Republic
Institute of Microbiology CAS v v i Vídeňská 1083 142 20 Praha 4 Czech Republic
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