Premium
TssA from Burkholderia cenocepacia : expression, purification, crystallization and crystallographic analysis
Author(s) -
Owen Hayley J.,
Sun Ruyue,
Ahmad Asma,
Sedelnikova Svetlana E.,
Baker Patrick J.,
Thomas Mark S.,
Rice David W.
Publication year - 2018
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/s2053230x18009706
Subject(s) - burkholderia cenocepacia , clade , protein subunit , phylogenetic tree , biology , crystallization , computational biology , gene , chemistry , burkholderia , genetics , bacteria , organic chemistry
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 D 16 symmetry, whereas the N‐terminal domain is not involved in subunit assocation.