TssA from Burkholderia cenocepacia: expression, purification, crystallization and crystallographic analysis

Acta Crystallogr F Struct Biol Commun. 2018 Sep 1;74(Pt 9):536-542. doi: 10.1107/S2053230X18009706. Epub 2018 Aug 29.

Abstract

TssA is a core component of the type VI secretion system, and phylogenetic analysis of TssA subunits from different species has suggested that these proteins fall into three distinct clades. Whilst representatives of two clades, TssA1 and TssA2, have been the subjects of investigation, no members of the third clade (TssA3) have been studied. Constructs of TssA from Burkholderia cenocepacia, a representative of clade 3, were expressed, purified and subjected to crystallization trials. Data were collected from crystals of constructs of the N-terminal and C-terminal domains. Analysis of the data from the crystals of these constructs and preliminary structure determination indicates that the C-terminal domain forms an assembly of 32 subunits in D16 symmetry, whereas the N-terminal domain is not involved in subunit assocation.

Keywords: Burkholderia cenocepacia; TssA; type VI secretion system.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Burkholderia cenocepacia / chemistry*
  • Burkholderia cenocepacia / classification
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Electrons*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • Membrane Proteins / chemistry*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Phylogeny
  • Protein Conformation, alpha-Helical
  • Protein Domains
  • Protein Subunits / chemistry*
  • Protein Subunits / genetics
  • Protein Subunits / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Type VI Secretion Systems / chemistry*

Substances

  • Bacterial Proteins
  • Membrane Proteins
  • Protein Subunits
  • Recombinant Proteins
  • Type VI Secretion Systems

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