Recombinant expression, purification, crystallization and preliminary X-ray diffraction analysis of the C-terminal DUF490(963-1138) domain of TamB from Escherichia coli

Acta Crystallogr F Struct Biol Commun. 2014 Sep;70(Pt 9):1272-5. doi: 10.1107/S2053230X14017403. Epub 2014 Aug 27.

Abstract

TamB is a recently described inner membrane protein that, together with its partner protein TamA, is required for the efficient secretion of a subset of autotransporter proteins in Gram-negative bacteria. In this study, the C-terminal DUF490963-1138 domain of TamB was overexpressed in Escherichia coli K-12, purified and crystallized using the sitting-drop vapour-diffusion method. The crystals belonged to the primitive trigonal space group P3121, with unit-cell parameters a = b = 57.34, c = 220.74 Å, and diffracted to 2.1 Å resolution. Preliminary secondary-structure and X-ray diffraction analyses are reported. Two molecules are predicted to be present in the asymmetric unit. Experimental phasing using selenomethionine-labelled protein will be undertaken in the future.

Keywords: DUF490; Escherichia coli; TamB; autotransporter.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Crystallization
  • Crystallography, X-Ray / methods*
  • DNA Primers
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / isolation & purification
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Protein Structure, Secondary
  • Recombinant Proteins / chemistry

Substances

  • DNA Primers
  • Escherichia coli Proteins
  • Recombinant Proteins