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Full‐length nisin immunity protein NisI from Lactococcus lactis in a lipid‐free form: crystallization and X‐ray analysis
Author(s) -
Jeong Jin Hee,
Ha Sung Chul
Publication year - 2017
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x17008214
Subject(s) - nisin , crystallization , crystallography , materials science , chemistry , biology , microbiology and biotechnology , organic chemistry , antimicrobial
NisI is a lantibiotic‐binding lipoprotein that is specific for nisin. Nisin‐producing microorganisms use NisI as an immunity protein for self‐protection against nisin. Here, the purification, crystallization and preliminary X‐ray diffraction of full‐length NisI from Lactobacillus lactis in a lipid‐free form (NisI 22‐C ) are reported. Importantly, reductive methylation of the lysine residues in NisI 22‐C was essential for initial crystallization. Only methylated NisI 22‐C crystallized. The optimized crystals of methylated NisI 22‐C were grown in 30–40 m M ammonium sulfate, 0.1 M sodium acetate pH 4.6, 16–18% PEG 4000 at 295 K and diffracted to 1.9 Å resolution. The crystal belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 45.99, b = 76.67, c = 76.39 Å, α = β = γ = 90.0°. Assuming the presence of one molecule in the asymmetric unit, the estimated Matthews coefficient ( V M ) is 2.58 Å 3 Da −1 and the estimated solvent content is 52.3%.