title: Overexpression, purification, crystallization and preliminary X-ray diffraction of the nisin resistance protein from Streptococcus agalactiae.
abstract: Nisin is a 34-amino-acid antimicrobial peptide produced by Lactococcus lactis belonging to the class of lantibiotics. Nisin displays a high bactericidal activity against various Gram-positive bacteria, including some human-pathogenic strains. However, there are some nisin-non-producing strains that are naturally resistant owing to the presence of the nsr gene within their genome. The encoded protein, NSR, cleaves off the last six amino acids of nisin, thereby reducing its bactericidal efficacy. An expression and purification protocol has been established for the NSR protein from Streptococcus agalactiae COH1. The protein was successfully crystallized using the vapour-diffusion method in hanging and sitting drops, resulting in crystals that diffracted X-rays to 2.8 and 2.2 Å, respectively.
authors.affiliation: Institute of Biochemistry, Heinrich-Heine-University, 40225 Düsseldorf, Germany.
authors.forename: Sakshi
authors.initials: S
authors.lastname: Khosa
authors.affiliation: Crystal Farm and X-ray Facility, Heinrich-Heine-University, 40225 Düsseldorf, Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Hoeppner
authors.affiliation: Protein Production Facility, Heinrich-Heine-University, 40225 Düsseldorf, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Kleinschrodt
authors.affiliation: Institute of Biochemistry, Heinrich-Heine-University, 40225 Düsseldorf, Germany.
authors.forename: Sander H J
authors.initials: SH
authors.lastname: Smits
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D009561:Nisin / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013292:Streptococcus agalactiae / Q000737:chemistry* / Q000201:enzymology; D014961:X-Ray Diffraction
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D009561:Nisin; D006867:Hydrolases
keywords: Lactococcus lactis; immunity; lantibiotic; lipoprotein; nisin; resistance