title: Cysteine-22 and cysteine-38 are not essential for the functions of maltoporin (LamB protein).
abstract: Maltoporin in the outer membrane of Escherichia coli contains two cysteine residues, at positions 22 and 38 in the primary sequence. The role of these residues in determining structural stability, and their contributions to the maltoporin binding sites for maltodextrins and bacteriophage lambda, was investigated. Site-directed mutagenesis was used to alter each of these residues to a serine. A double mutant lacking both cysteines was also isolated. None of the substitutions affected maltodextrin binding or the binding of phage lambda, suggesting the variant proteins retain a native binding-site conformation. The mutants were assembled at wild-type levels into the outer membrane as maltoporin trimers but the temperature-stability of the trimer greater than monomer dissociation was slightly reduced in the presence of the Cys 38 substitution. However, it is unlikely that the stability of trimers was due to disulfide linkages between subunits since the native trimers are stable under highly reducing conditions in the presence of SDS; more likely the Cys greater than Ser substitutions slightly perturb intra- or inter-subunit hydrophobic interactions in regions predicted to span across the outer membrane.
authors.affiliation: Department of Microbiology, University of Sydney, Australia.
authors.forename: T
authors.initials: T
authors.lastname: Ferenci
authors.forename: S
authors.initials: S
authors.lastname: Stretton
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D003545:Cysteine / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology; D009154:Mutation; D018272:Porins; D011487:Protein Conformation; D011991:Receptors, Virus / Q000235:genetics / Q000502:physiology*; D013329:Structure-Activity Relationship
chemical_list: D001425:Bacterial Outer Membrane Proteins; D018272:Porins; D011991:Receptors, Virus; C016532:maltoporins; D003545:Cysteine