Purification and chemical characterization of type 1 pili isolated from Klebsiella pneumoniae

R. C. Fader, L. K. Duffy, C. P. Davis, A. Kurosky

    Research output: Contribution to journalArticle

    25 Citations (Scopus)

    Abstract

    Type 1 pili were purified from a Klebsiella pneumoniae strain isolated from a human urinary tract infection. The pili were removed from the bacteria by mechanical shearing, precipitated out of solution by ammonium sulfate, solubilized in a deoxycholate-containing buffer, and finally purified by gel filtration. Chemical characterization of the isolated pili revealed a single protein subunit (pilin) which had a M(r) = 21,500. Amino acid compositional analysis revealed a high content of residues that contribute significantly to secondary structure. Automated sequence analysis of the NH2-terminal region revealed a striking homology (79% identity) with type 1 pili of Escherichia coli. In contrast, NH2-terminal sequence comparison of K. pneumoniae pilin with other previously reported bacterial pilins showed no significant homology. No immunological cross-reactivity was detectable between E. coli and K. pneumoniae pili when tested by Ouchterlony double immunodiffusion or by rocket immunoelectrophoresis. The results of this study, when compared to other studies of bacterial pili, indicate that type 1 pili from members of the Enterobacteriaceae share morphological similarities and that their monomeric subunits are chemically similar. In addition, these results give strong evidence that the type 1 pilins of the enteric bacteria represent a separate class of homologous pilins.

    Original languageEnglish (US)
    Pages (from-to)3301-3305
    Number of pages5
    JournalJournal of Biological Chemistry
    Volume257
    Issue number6
    StatePublished - 1982

    Fingerprint

    Fimbriae Proteins
    Klebsiella pneumoniae
    Purification
    Enterobacteriaceae
    Escherichia coli
    Bacteria
    Bacterial Fimbriae
    Immunoelectrophoresis
    Deoxycholic Acid
    Immunodiffusion
    Protein Subunits
    Ammonium Sulfate
    Rockets
    Shearing
    Urinary Tract Infections
    Gel Chromatography
    Sequence Analysis
    Buffers
    Gels
    Amino Acids

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Fader, R. C., Duffy, L. K., Davis, C. P., & Kurosky, A. (1982). Purification and chemical characterization of type 1 pili isolated from Klebsiella pneumoniae. Journal of Biological Chemistry, 257(6), 3301-3305.

    Purification and chemical characterization of type 1 pili isolated from Klebsiella pneumoniae. / Fader, R. C.; Duffy, L. K.; Davis, C. P.; Kurosky, A.

    In: Journal of Biological Chemistry, Vol. 257, No. 6, 1982, p. 3301-3305.

    Research output: Contribution to journalArticle

    Fader, RC, Duffy, LK, Davis, CP & Kurosky, A 1982, 'Purification and chemical characterization of type 1 pili isolated from Klebsiella pneumoniae', Journal of Biological Chemistry, vol. 257, no. 6, pp. 3301-3305.
    Fader, R. C. ; Duffy, L. K. ; Davis, C. P. ; Kurosky, A. / Purification and chemical characterization of type 1 pili isolated from Klebsiella pneumoniae. In: Journal of Biological Chemistry. 1982 ; Vol. 257, No. 6. pp. 3301-3305.
    @article{3db2cd4044364611bf565cd54e57bbd9,
    title = "Purification and chemical characterization of type 1 pili isolated from Klebsiella pneumoniae",
    abstract = "Type 1 pili were purified from a Klebsiella pneumoniae strain isolated from a human urinary tract infection. The pili were removed from the bacteria by mechanical shearing, precipitated out of solution by ammonium sulfate, solubilized in a deoxycholate-containing buffer, and finally purified by gel filtration. Chemical characterization of the isolated pili revealed a single protein subunit (pilin) which had a M(r) = 21,500. Amino acid compositional analysis revealed a high content of residues that contribute significantly to secondary structure. Automated sequence analysis of the NH2-terminal region revealed a striking homology (79{\%} identity) with type 1 pili of Escherichia coli. In contrast, NH2-terminal sequence comparison of K. pneumoniae pilin with other previously reported bacterial pilins showed no significant homology. No immunological cross-reactivity was detectable between E. coli and K. pneumoniae pili when tested by Ouchterlony double immunodiffusion or by rocket immunoelectrophoresis. The results of this study, when compared to other studies of bacterial pili, indicate that type 1 pili from members of the Enterobacteriaceae share morphological similarities and that their monomeric subunits are chemically similar. In addition, these results give strong evidence that the type 1 pilins of the enteric bacteria represent a separate class of homologous pilins.",
    author = "Fader, {R. C.} and Duffy, {L. K.} and Davis, {C. P.} and A. Kurosky",
    year = "1982",
    language = "English (US)",
    volume = "257",
    pages = "3301--3305",
    journal = "Journal of Biological Chemistry",
    issn = "0021-9258",
    publisher = "American Society for Biochemistry and Molecular Biology Inc.",
    number = "6",

    }

    TY - JOUR

    T1 - Purification and chemical characterization of type 1 pili isolated from Klebsiella pneumoniae

    AU - Fader, R. C.

    AU - Duffy, L. K.

    AU - Davis, C. P.

    AU - Kurosky, A.

    PY - 1982

    Y1 - 1982

    N2 - Type 1 pili were purified from a Klebsiella pneumoniae strain isolated from a human urinary tract infection. The pili were removed from the bacteria by mechanical shearing, precipitated out of solution by ammonium sulfate, solubilized in a deoxycholate-containing buffer, and finally purified by gel filtration. Chemical characterization of the isolated pili revealed a single protein subunit (pilin) which had a M(r) = 21,500. Amino acid compositional analysis revealed a high content of residues that contribute significantly to secondary structure. Automated sequence analysis of the NH2-terminal region revealed a striking homology (79% identity) with type 1 pili of Escherichia coli. In contrast, NH2-terminal sequence comparison of K. pneumoniae pilin with other previously reported bacterial pilins showed no significant homology. No immunological cross-reactivity was detectable between E. coli and K. pneumoniae pili when tested by Ouchterlony double immunodiffusion or by rocket immunoelectrophoresis. The results of this study, when compared to other studies of bacterial pili, indicate that type 1 pili from members of the Enterobacteriaceae share morphological similarities and that their monomeric subunits are chemically similar. In addition, these results give strong evidence that the type 1 pilins of the enteric bacteria represent a separate class of homologous pilins.

    AB - Type 1 pili were purified from a Klebsiella pneumoniae strain isolated from a human urinary tract infection. The pili were removed from the bacteria by mechanical shearing, precipitated out of solution by ammonium sulfate, solubilized in a deoxycholate-containing buffer, and finally purified by gel filtration. Chemical characterization of the isolated pili revealed a single protein subunit (pilin) which had a M(r) = 21,500. Amino acid compositional analysis revealed a high content of residues that contribute significantly to secondary structure. Automated sequence analysis of the NH2-terminal region revealed a striking homology (79% identity) with type 1 pili of Escherichia coli. In contrast, NH2-terminal sequence comparison of K. pneumoniae pilin with other previously reported bacterial pilins showed no significant homology. No immunological cross-reactivity was detectable between E. coli and K. pneumoniae pili when tested by Ouchterlony double immunodiffusion or by rocket immunoelectrophoresis. The results of this study, when compared to other studies of bacterial pili, indicate that type 1 pili from members of the Enterobacteriaceae share morphological similarities and that their monomeric subunits are chemically similar. In addition, these results give strong evidence that the type 1 pilins of the enteric bacteria represent a separate class of homologous pilins.

    UR - http://www.scopus.com/inward/record.url?scp=0020025301&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0020025301&partnerID=8YFLogxK

    M3 - Article

    C2 - 6120941

    AN - SCOPUS:0020025301

    VL - 257

    SP - 3301

    EP - 3305

    JO - Journal of Biological Chemistry

    JF - Journal of Biological Chemistry

    SN - 0021-9258

    IS - 6

    ER -