Open AccessSignal Sequence Mutations as Tools for the Characterization of LamB Folding Intermediates
Author(s) -
Amy R. Duguay,
Thomas J. Silhavy
Publication year - 2002
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.184.24.6918-6928.2002
Subject(s) - biology , trimer , mutant , folding (dsp implementation) , amino acid , sequence (biology) , mutation , peptide sequence , signal peptide , biochemistry , biophysics , chemistry , gene , dimer , organic chemistry , electrical engineering , engineering
lamBA23DA25Y and lamBA23YA25Y tether LamB to the inner membrane by blocking signal sequence processing. We isolated suppressors of lamBA23DA25Y and lamBA23YA25Y, all of which mapped within the LamB signal sequence. Most interesting were mutations that changed an amino acid with a strong positive charge to an amino acid with no charge. Further characterization of two such suppressors revealed that they produce functional LamB that is localized to the outer membrane with its entire signal sequence still attached. Biochemical analysis shows that mutant LamB monomer chases into an oligomeric species with properties different from those of wild-type LamB trimer. Because assembly of mutant LamB is slowed, these mutations provide useful tools for the characterization of LamB folding intermediates.