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 Mr = 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.
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002458:Cell Fractionation; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000648:ultrastructure*; D007711:Klebsiella pneumoniae / Q000648:ultrastructure*; D008565:Membrane Proteins / Q000302:isolation & purification*; D008854:Microscopy, Electron; D008970:Molecular Weight; D013045:Species Specificity
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D008565:Membrane Proteins; D036781:Fimbriae Proteins