Skip to main content
SpringerLink
Log in

Pullulanase: Model protein substrate for the general secretory pathway of gram-negative bacteria

  • Papers
  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Pullulanase ofKlebsiella oxytoca is one of a wide variety of extracellular proteins that are secreted by Gram-negative bacteria by the complex main terminal branch (MTB) of the general secretory pathway. The roles of some of the 14 components of the MTB are now becoming clear. In this review it is proposed that most of these proteins form a complex, the secreton, that spans the cell envelope to control the opening and closing of channel in the outer membrane. Progress toward the goal of testing this model is reviewed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Akrim M., Bally M., Ball G., Tommassen J., Teerink H., Filloux A., Lazdunski A.: Xcp-mediated protein secretion inPseudomonas aeruginosa: identification of two additional genes and evidence for regulation ofxcp gene expression.Mol. Microbiol. 10, 431–443 (1993).

    PubMed  CAS  Google Scholar 

  • Andrews S.C., Harrison P.M., Guest J.R.: Cloning, sequencing and mapping of the bacterioferritin gene (bfr) ofEscherichia coli K-12.J. Bacteriol. 171, 3940–3947 (1989).

    PubMed  CAS  Google Scholar 

  • Bally M., Ball G., Badère A., Lazdunski A.: Protein secretion inPseudomonas aeruginosa: thexcpA gene encodes an integral membrane protein homologous toKlebsiella pneumoniae secretion function PulO.J. Bacteriol 173, 479–486 (1991).

    PubMed  CAS  Google Scholar 

  • Bally M., Filloux A., Akrim M., Ball G., Lazdunski A., Tommassen J.: Protein secretion inPseudomonas aeruginosa: characterization of sevenxcp genes and processing of secretory apparatus components by prepilin peptidase.Mol. Microbiol. 6, 1121–1131 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Bortoli-German I., Brun E., Py B., Chippaux M., Barras F.: Periplasmic disulphide bond formation is essential for cellulase secretion by the plant pathogenErwinia chrysanthemi.Mol. Microbiol. 11, 545–553 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Condemine G., Dorel C., Hugouvieux-Cotte-Pattat N., Robert-Baudouy J.: Some of theout genes involved in the secretion of pectate lyases inErwinia chrysanthemi are regulated bykdgR.Mol. Microbiol. 6, 3199–3211 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Drake S.L., Koomey M.: The product of thepilQ gene is essential for biogenesis of type IV pili inNeisseria gonorrhoeae.Mol. Microbiol. 18, 975–986 (1995).

    Article  PubMed  CAS  Google Scholar 

  • Dums F., Dow J.M., Daniels M.J.: Structural characterization of protein secretion genes of the bacterial phytopathogenXanthomonas campestris pathovarcampestris: relatedness to secretion systems of other Gram-negative bacteria.Mol. Gen. Genet. 229, 357–364 (1991).

    Article  PubMed  CAS  Google Scholar 

  • D'Enfert C., Pugsley A.P.:Klebsiella pneumoniae pulS gene encodes an outer membrane lipoprotein required for pullulanase secretion.J. Bacteriol. 171, 3673–3679 (1989).

    PubMed  Google Scholar 

  • D'Enfert C., Reyss I., Wandersman C., Pugsley A.P.: Protein secretion by Gram-negative bacteria: characterization of two membrane proteins required for pullulanase secretion byEscherichia coli K-12.J. Biol. Chem. 264, 17462–17468 (1989).

    PubMed  Google Scholar 

  • D'Enfert C., Ryter A., Pugsley A.P.: Cloning and expression inEscherichia coli of theKlebsiella pneumoniae genes for production, surface localization and secretion of the lipoprotein pullulanase.EMBO J. 6, 3531–3538 (1987).

    PubMed  Google Scholar 

  • Francetic O., Pugsley A.P.: The cryptic general secretory pathway (gsp) operon ofEscherichia coli K-12 encodes functional proteins.J. Bacteriol. 178, 3544–3549 (1996).

    PubMed  CAS  Google Scholar 

  • Genin S., Boucher C.A.: A superfamily of proteins involved in different secretion pathways in Gram-negative bacteria: modular structure and specificity of the N-terminal domain.Mol. Gen. Genet. 243, 112–118 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Hardie K.R., Schulze A., Parker M.W., Buckley J.T.:Vibrio spp. secrete proaerolysin as a folded dimer without the need for disulphide bond formation.Mol. Microbiol. 17, 1035–1044 (1995).

    Article  PubMed  CAS  Google Scholar 

  • Hardie K.R., Lory S., Pugsley A.P.: Insertion of an outer membrane protein inEscherichia coli requires a chaperone-like protein.EMBO J. 15, 978–988 (1996a).

    PubMed  CAS  Google Scholar 

  • Hardie K.R., Seydel A., Guilvout I., Pugsley A.P.: The secretin-specific chaperone protein of the general secretory pathway: separation of proteolytic protection, and piloting functions.Mol. Microbiol. 22, 967–976 (1996b).

    Article  PubMed  CAS  Google Scholar 

  • He S.Y., Lindeberg M., Chatterjee A.K., Collmer A.: ClonedErwinia chrysanthemi out genes enableEscherichia coli to selectively secrete a diverse family of heterologous proteins into its milieu.Proc. Nat. Acad. Sci. USA 88, 1079–1083 (1991).

    Article  PubMed  CAS  Google Scholar 

  • Hirst T.R., Holmgren J.: Conformation of protein secreted across bacterial outer membanes: a study of enterotoxin translocation fromVibrio cholerae.Proc. Nat. Acad. Sci. USA 84, 7418–7422 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Hobbs M., Mattick J.S.: Common components in the assembly of type 4 fimbriae, DNA transfer systems, filamentous phage and protein-secretion apparatus: a general system for the formation of surface-associated protein complexes.Mol. Microbiol. 10, 233–243 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Howard S.P., Critch J., Bedi A.: Isolation and analysis of eightexe genes and their involvement in extracellular protein secretion and outer membrane assembly inAeromonas hydrophila.J. Bacteriol. 175, 6695–6703 (1993).

    PubMed  CAS  Google Scholar 

  • Hu N.-T., Hung M.-I., Chiou S.-J., Tang F., Chiang D.-C., Huang H.-Y., Wu C.-Y.: Cloning and characterization of a gene required for the secretion of extracellular enzymes across the outer membrane byXanthomonas campestris.J. Bacteriol. 174, 2679–2687 (1992).

    PubMed  CAS  Google Scholar 

  • Hu N.-t., Hung M.-N., Liao C.-T., Lin M.-H.: Subcellular location of XpsD, a protein required for extracellular protein secretion byXanthomonas campestris pv.campestris.Microbiology 141, 1395–1406 (1995).

    Article  PubMed  CAS  Google Scholar 

  • Jahagirdar R., Howard S.P.: Isolation and characterization of a secondexe operon required for extracellular protein secretion inAeromonas hydrophila.J. Bacteriol. 176, 6819–6826 (1994).

    PubMed  CAS  Google Scholar 

  • Jiang B., Howard S.P.: TheAeromonas hydrophila exeE gene required both for protein secretion and normal outer membrane biogenesis, is a member of a general secretion pathway.Mol. Microbiol. 6, 1351–1361 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Kornacker M.G., Pugsley A.P.: Molecular characterization ofpulA and its product, pullulanase, a secreted enzyme ofKlebsiella pneumoniae UNF5023.Mol. Microbiol. 4, 73–85 (1989).

    Article  Google Scholar 

  • Kornacker M.G., Pugsley A.P.: The normally periplasmic enzyme β-lactamase is specifically and efficiently translocated through theEscherichia coli outer membrane when it is fused to the cell surface enzyme pullulanase.Mol. Microbiol. 4, 1101–1109 (1990).

    Article  PubMed  CAS  Google Scholar 

  • Lindeberg M., Salmond G.P.C., Collmer A.: Complementation of deletion mutations in a cloned functional cluster ofErwinia chrysanthemi out genes withErwinia carotovora out homologs reveals OutC and OutD as candidate gatekeepers of species-specific secretion of proteinsvia the Type II pathway.Mol. Microbiol. 20, 175–190 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Linderoth N.A., Model P., Russel M.: Essential role of a sodium dodecyl sulfate-resistant protein IV multimer in assembly-export of filamentous phage.J. Bacteriol. 178, 1962–1970 (1996).

    PubMed  CAS  Google Scholar 

  • Michaelis S., Chapon C., D'Enfert C., Pugsley A.P., Schwartz M.: Characterization and expression of the structural gene for pullulanase, a maltose-inducible secreted protein ofKlebsiella pneumoniae.J. Bacteriol. 164, 633–638 (1985).

    PubMed  CAS  Google Scholar 

  • Nunn D., Bergman S., Lory S.: Products of three accessory genes,pilB, pilC, andpilD, are required for biogenesis ofPseudomonas aeruginosa pili.J. Bacteriol. 172, 2911–2919 (1990).

    PubMed  CAS  Google Scholar 

  • Nunn D., Lory S.: Components of the protein excretion apparatus ofPseudomonas aeruginosa are processed by the type IV prepilin peptidase.Proc. Nat. Acad. Sci. USA 89, 47–51 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Possot O., D'Enfert C., Reyss I., Pugsley A.P.: Pullulanase secretion inEscherichia coli K12 requires a cytoplasmic protein and a putative polytopic cytoplasmic membrane protein.Mol. Microbiol. 6, 95–105 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Possot, O., Pugsley A.P.: The conserved tetracysteine motif in the general secretory pathway component PulE is required for efficient pullulanase secretion.Gene, in press (1997).

  • Possot O., Pugsley A.P.: Molecular characterization of PulE, a protein required for pullulanase secretion.Mol. Microbiol. 12, 287–299 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Pugsley A.P.: Translocation of a folded protein across the outer membranevia the general secretory pathway inEscherichia coli.Proc. Nat. Acad. Sci. USA 89, 12058–12062 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Pugsley A.P.: The complete general secretory pathway in gram-negative bacteria.Microbiol. Rev. 57, 50–108 (1993a).

    PubMed  CAS  Google Scholar 

  • Pugsley A.P.: Processing and methylation of PulG, a pilin-like component of the general secretory pathway ofKlebsiella oxytoca.Mol. Microbol. 9, 295–308 (1993b).

    Article  CAS  Google Scholar 

  • Pugsley A.P.: Multimers of the precursor of a type IV pilin-like component of the general secretory pathway are unrelated to pili.Mol. Microbiol. 20, 1235–1245 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Pugsley A.P., Chapon C., Schwartz M.: Extracellular pullulanase ofKlebsiella pneumoniae is a lipoprotein.J. Bacteriol. 166, 1083–1088 (1986).

    PubMed  CAS  Google Scholar 

  • Pugsley A.P., Dupuy B.: An enzyme with type IV prepilin peptidase activity is required to process a component of the general extracellular protein secretion pathway ofKlebsiella oxytoca.Mol. Microbiol. 6, 751–760 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Pugsley A.P., Kornacker M.G., Poquet I.: The general secretion pathway is directly required for extracellular pullulanase secretion inEscherichia coli.Mol. Microbiol. 5, 343–352 (1991a).

    Article  PubMed  CAS  Google Scholar 

  • Pugsley A.P., Poquet I., Kornacker M.G.: Two distinct steps in pullulanase secretion byEscherichia coli K12.Mol. Microbiol. 5, 865–873 (1991b).

    Article  PubMed  CAS  Google Scholar 

  • Pugsley A.P., Possot O.: The general secretory pathway ofKlebsiella oxytoca: no evidence for relocalization or assembly of pilin-like PulG protein into a multiprotein complex.Mol. Microbiol. 10, 665–674 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Pugsley A.P., Reyss I.: Five genes at the 3′ end of theKlebsiella pneumoniae pulC operon are required for pullulanase secretion.Mol. Microbiol. 4, 365–379 (1990).

    Article  PubMed  CAS  Google Scholar 

  • Reyss I., Pugsley A.P.: Five additional genes in thepulC-O operon of the Gram-negative bacteriumKlebsiella oxytoca UNF5023 that are required for pullulanase secretion.Mol. Gen. Genet. 222, 176–184 (1990).

    Article  PubMed  CAS  Google Scholar 

  • Russel M.: Mutants at conserved positions in gene IV, a gene required for assembly and secretion of filamentous phages.Mol. Microbiol. 14, 357–369 (1994a).

    Article  PubMed  CAS  Google Scholar 

  • Russel M.: Phage assembly: a paradigm for bacterial virulence factor export?Science 265, 612–614 (1994b).

    Article  PubMed  CAS  Google Scholar 

  • Russel M., Kazmierczak B.: Analysis of the structure and subcellular location of filamentous phage pIV.J. Bacteriol. 175, 3998–4007 (1993).

    PubMed  CAS  Google Scholar 

  • Sandkvist M., Bagdasarian M., Howard S.P., DiRita V.J.: Interaction between the autokinase EpsE and EpsL in the cytoplasmic membrane is required for extracellular secretion inVibrio cholerae.EMBO J. 14, 1664–1673 (1995).

    PubMed  CAS  Google Scholar 

  • Sauvonnet N., Poquet I., Pugsley A.P.: Extracellular secretion of pullulanase is unaffected by minor sequence changes but is usually prevented by adding reporter proteins to its N-or C-terminal end.J. Bacteriol. 177, 5238–5246 (1995).

    PubMed  CAS  Google Scholar 

  • Sauvonnet N., Pugsley A.P.: Identification of two regions ofKlebsiella oxytoca pullulanase that together are capable of promoting β-lactamase secretion by the general secretory pathway.Mol. Microbiol. 22, 1–7 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Skare J.T., Ahmer B.M.M., Seachord C.L., Darveau R.P., Postle K.: Energy transduction between membranes. TonB, a cytoplasmic membrane protein, can be chemically cross-linkedin vivo to the outer membrane receptor FepA.J. Biol. Chem. 268, 16302–16308 (1993).

    PubMed  CAS  Google Scholar 

  • Strom M.S., Lory S.: Kinetics and sequence specificity of processing of prepilin by PilD, the type IV leader peptidase ofPseudomonas aeruginosa.J. Bacteriol. 174, 7345–7351 (1992).

    PubMed  CAS  Google Scholar 

  • Strom M.S., Bergman P., Lory S.: Identification of active-site cysteines in the conserved domain of PilD, the bifunctional type IV pilin leader peptidase/N-methyltransferase ofPseudomonas aeruginosa.J. Biol. Chem. 268, 15788–15794 (1993a).

    PubMed  CAS  Google Scholar 

  • Strom M.S., Nunn D.N., Lory S.: A single bifunctional enzyme, PilD, catalyzes cleavage and N-methylation of proteins belonging to the type IV pilin family.Proc. Nat. Acad. Sci. USA 90, 2404–2408 (1993b).

    Article  PubMed  CAS  Google Scholar 

  • Turner L.R., Cano-Lara J., Nunn D.N., Lory S.: Mutations in the consensus ATP-binding sites of XcpR and PilB eliminate extracellular protein secretion and pilus biogenesis inPseudomonas aeruginosa.J. Bacteriol. 175, 4962–4969 (1993).

    PubMed  CAS  Google Scholar 

  • Vidal-Ingigliardi D., Richet E., Raibaud O.: Two MalT binding sites in direct repeat. A structural motif involved in the activation of all of the promoters of the maltose regulons inEscherihia coli andKlebsiella pneumoniae.J. Mol. Biol. 218, 323–334 (1991).

    Article  PubMed  CAS  Google Scholar 

  • Wong K.R., Buckley T.J.: Proton motive force involved in protein transport across the outer membrane ofAeromonas salmonicida.Science 246, 654–656 (1989).

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Unité de Génétique Moléculaire, CNRS URA 1149, Institut Pasteur, 75724, Paris Cedex 15, France

    A. P. Pugsley, O. Francetic, K. Hardie, O. M. Possot, N. Sauvonnet & A. Seydel

Authors
  1. A. P. Pugsley
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. O. Francetic
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. Hardie
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. O. M. Possot
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. N. Sauvonnet
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. Seydel
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pugsley, A.P., Francetic, O., Hardie, K. et al. Pullulanase: Model protein substrate for the general secretory pathway of gram-negative bacteria. Folia Microbiol 42, 184–192 (1997). https://doi.org/10.1007/BF02818976

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02818976

Keywords

Search

Navigation