Purification, crystallization and preliminary X-ray crystallographic analysis of the outer membrane lipoprotein NlpE from Escherichia coli
- Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502 (Japan)
- Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan)
A water-soluble mutant of the outer membrane lipoprotein NlpE has been overexpressed, purified and crystallized. Diffraction data from two crystal forms obtained under two different conditions were collected to 2.8 and 3.0 Å resolution and processed in space groups P4{sub 3}2{sub 1}2 and C2, respectively. The outer membrane lipoprotein NlpE functions in stress response by activating the Cpx signal transduction pathway. The nonlipidated Cys1Ala mutant of NlpE with a C-terminal His tag from Escherichia coli was constructed, overexpressed and purified. Crystals of NlpE were grown in two distinct forms by the sitting-drop vapour-diffusion method at 298 K. The tetragonal crystals diffracted to 2.8 Å resolution and belong to space group P4{sub 3}2{sub 1}2. The monoclinic crystals diffracted to 3.0 Å resolution and belong to space group C2. Initial phases were obtained from a tetragonal crystal of selenomethionylated protein by the MAD method.
- OSTI ID:
- 22356426
- Journal Information:
- Acta Crystallographica. Section F, Vol. 62, Issue Pt 12; Other Information: PMCID: PMC2225377; PMID: 17142903; PUBLISHER-ID: pu5165; OAI: oai:pubmedcentral.nih.gov:2225377; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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