Solution NMR Structure of TolAIII Bound to a Peptide Derived from the N-terminus of TolB
The lipoprotein Pal stabilises the bacterial outer membrane during constriction by a mobilisation-and-capture mechanism.
Peptide sequence represents residues 22 to 34 of the TolB sequence. This is the region of TolB that binds to TolAIII.
This is the 3rd domain of the TolA protein.
Cell envelope integrity/translocation protein TolA
HMRALAELLSDTTERQQALADEVGSEVTGSLDDLIVNLVSQQWRRPPSARNGMSVEVLIEMLPDGTITNASVSRSSGDKP FDSSAVAAVRNVGRIPEMQQLPRATFDSLYRQRRIIFKPEDLSL
Biotechnology and Biological Sciences Research Council
8 rounds of iterative structure calculation using ARIA protocol
20 lowest energy structures further refined using MD in water via ARIA protocol
0.6 mM [U-98% 15N] TolAIII, 3 mM TolBp, 20 mM sodium phosphate, 60 mM sodium chloride, 95% H2O/5% D2O
0.6 mM [U-98% 13C; U-98% 15N] TolAIII, 3 mM TolBp, 20 mM sodium phosphate, 60 mM sodium chloride, 95% H2O/5% D2O
1.1 mM TolAIII, 0.2 mM [U-98% 13C; U-98% 15N] TolBp, 20 mM sodium phosphate, 60 mM sodium chloride, 95% H2O/5% D2O
0.6 mM [U-98% 13C; U-98% 15N] TolAIII, 3 mM TolBp, 20 mM sodium phosphate, 60 mM sodium chloride, 100% D2O
0.6 mM [U-98% 15N] TolAIII, 3 mM TolBp, 20 mM sodium phosphate, 60 mM sodium chloride, 5 % C12E6/n-hexanol, 95% H2O/5% D2O
Used for measurement of residual dipolar couplings. Alignment medium is mixture of C12E6 and n-hexanol as described by Ruckert & Otting (2000).
Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
Brunger, Adams, Clore, Gros, Nilges and Read
The 1:1 stoichiometry was determined by ITC. In addition, NMR titrations confirmed this.
TolA, TolB, Pal, bacterial outer membrane, PROTEIN BINDING
RALAELLSDTTERQQALADEVGSEVTGSLDDLIVNLVSQQWRRPPSARNGMSVEVLIEMLPDGTITNASVSRSSGDKPFD SSAVAAVRNVGRIPEMQQLPRATFDSLYRQRRIIFKPEDLSL