The Escherichia coli RnlA-RnlB toxin-antitoxin complex

Subtitle:production, characterization and crystallization

The Escherichia coli rnlAB operon encodes a toxin-antitoxin module that is involved in protection against infection by bacteriophage T4. The full-length RnlA-RnlB toxin-antitoxin complex as well as the toxin RnlA were purified to homogeneity and crystallized. When the affinity tag is placed on RnlA, RnlB is largely lost during purification and the resulting crystals exclusively comprise RnlA. A homogeneous preparation of RnlA-RnlB containing stoichiometric amounts of both proteins could only be obtained using a His tag placed C-terminal to RnlB. Native mass spectrometry and SAXS indicate a 1:1 stoichiometry for this RnlA-RnlB complex. Crystals of the RnlA-RnlB complex belonged to space group C2, with unit- cell parameters a = 243.32, b = 133.58, c = 55.64 angstrom, beta = 95.11 degrees, and diffracted to 2.6 angstrom resolution. The presence of both proteins in the crystals was confirmed and the asymmetric unit is likely to contain a heterotetramer with RnlA(2):RnlB(2) stoichiometry.

Publication year:2020

Keywords:A1 Journal article

BOF-keylabel:yes

BOF-publication weight:0.1

CSS-citation score:1

Authors:International

Authors from:Government

Accessibility:Closed