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

• Published in: Acta crystallographica. Section F, Structural biology communications Vol. 74; no. 9; pp. 536 - 542
• Main Authors: Owen, Hayley J., Sun, Ruyue, Ahmad, Asma, Sedelnikova, Svetlana E., Baker, Patrick J., Thomas, Mark S., Rice, David W.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.09.2018; Wiley Subscription Services, Inc
• Subjects: Amino Acid Sequence; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Burkholderia cenocepacia; Burkholderia cenocepacia - chemistry; Burkholderia cenocepacia - classification; Cloning, Molecular; Crystal structure; Crystallization; Crystallography; Crystallography, X-Ray; Crystals; Data processing; Domains; 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; Proteins; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Research Communications; Secretion; TssA; type VI secretion system; Type VI Secretion Systems - chemistry; type VI secretion system; TssA; Burkholderia cenocepacia
• ISSN: 2053-230X; 2053-230X
• DOI: 10.1107/S2053230X18009706

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. Analysis of the noncrystallographic symmetry of crystals of the C‐terminal domain of Burkholderia cenocepacia TssA indicates a quaternary structure of 32 subunits in D16 symmetry.