Overexpression, purification, crystallization and preliminary X-ray diffraction of the nisin resistance protein from Streptococcus agalactiae

Acta Crystallogr F Struct Biol Commun. 2015 Jun;71(Pt 6):671-5. doi: 10.1107/S2053230X15006226. Epub 2015 May 20.

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.

Keywords: Lactococcus lactis; immunity; lantibiotic; lipoprotein; nisin; resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Anti-Bacterial Agents / chemistry
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Drug Resistance, Bacterial
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Hydrolases / chemistry*
  • Hydrolases / genetics
  • Molecular Sequence Data
  • Nisin / chemistry
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Streptococcus agalactiae / chemistry*
  • Streptococcus agalactiae / enzymology
  • X-Ray Diffraction

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Recombinant Proteins
  • Nisin
  • Hydrolases