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Interactions among FliN, FliH and FliI of the Salmonella enterica flagellar export apparatus govern secretion
Author(s) -
Wilson Jo Leanna,
Scott Israel M.,
McMurry Jonathan L.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.664.3
Subject(s) - chemistry , affinities , secretion , surface plasmon resonance , biophysics , biochemistry , nanotechnology , biology , materials science , nanoparticle
Using two optical biosensing methods, surface plasmon resonance and biolayer interferometry, a study of the interactions among proteins involved in flagellar type III secretion was undertaken to understand the mechanism of substrate delivery to the membrane‐embedded export complex. An interaction between C‐ring protein FliN and export apparatus protein FliH has been proposed to be a mode of targeting substrates via a complex that is necessarily dynamic as the position of FliN at the base of the C‐ring is too far from the membrane to allow the complex to remain associated during export. Both techniques allow quantitative measurement of association and dissociation. Rate and affinity constants are reported for pairwise interactions; both FliN‐FliH and FliH‐FliI have submicromolar K D s. Experiments investigating the conditions necessary to dissociate FliN from FliH or FliH from FliI are described. Substrates were screened for binding to FliH and FliI in the presence and absence of FliN. Kinetic analysis will allow us to rank affinities, determine how binding changes in the presence of other participants, search for export signals within substrates etc., developing a better understanding of type III secretion. This work was supported by grants from the Public Health Service (R15GM080701), NSF (CHE 0922699), the Research Corporation (CC6900) and a KSU Mentor‐Protégé Grant (to J.L.W.).