title: The C-terminal domain of the secretin PulD contains the binding site for its cognate chaperone, PulS, and confers PulS dependence on pIVf1 function.
abstract: Related outer membrane proteins, termed secretins, participate in the secretion of macromolecules across the outer membrane of many Gram-negative bacteria. In the pullulanase-secretion system, PulS, an outer membrane-associated lipoprotein, is required both for the integrity and the proper outer membrane localization of the PulD secretin. Here we show that the PulS-binding site is located within the C-terminal 65 residues of PulD. Addition of this domain to the filamentous phage secretin, pIV, or to the unrelated maltose-binding protein rendered both proteins dependent on PulS for stability. A chimeric protein composed of bacteriophage f1 pIV and the C-terminal domain of PuID required properly localized PulS to support phage assembly. An in vivo complex formed between the pIV-PulD65 chimera and PulS was detected by co-immunoprecipitation and by affinity chromatography.
authors.affiliation: Rockefeller University, New York, New York 10021, USA.
authors.forename: S
authors.initials: S
authors.lastname: Daefler
authors.forename: I
authors.initials: I
authors.lastname: Guilvout
authors.forename: K R
authors.initials: KR
authors.lastname: Hardie
authors.forename: A P
authors.initials: AP
authors.lastname: Pugsley
authors.forename: M
authors.initials: M
authors.lastname: Russel
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis; D001435:Bacteriophages / Q000254:growth & development / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D006026:Glycoside Hydrolases / Q000378:metabolism; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000378:metabolism / Q000821:virology; D006360:Heat-Shock Proteins / Q000096:biosynthesis; D018832:Molecular Chaperones / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012633:Secretin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D017931:DNA Primers; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; C069816:phage shock protein, Bacteria; C061377:PulD protein, Klebsiella pneumoniae; D012633:Secretin; D006026:Glycoside Hydrolases; C100181:pullulanase