Comparative studies of the amino acid and nucleotide sequences of pilin derived from Pseudomonas aeruginosa PAK and PAO

Abstract

The entire amino acid sequence for Pseudomonas aeruginosa PAO pilin was determined through peptide sequencing and from the complete nucleotide sequence encoding the pilin gene. The precursor PAO pilin is 149 amino acids in length which includes a 6-amino-acid positively charged leader sequence. Comparison of the amino acid sequences of pilin produced by P. aeruginosa PAO and PAK reveals a region of high homology corresponding to the leader peptide and residues 1 to 54 of the mature pilin. The amino acid sequence of the peptide encompassing the major antigenic determinant of PAK differs greatly from that of the equivalent region in PAO. The C-terminal regions of these proteins are semiconserved. Few major differences were found

The pilE gene product of Pseudomonas aeruginosa, required for pilus biogenesis, shares amino acid sequence identity with the N‐termini of type 4 prepilin proteins

“…Antigenic analysis employing PilA peptides revealed that the protein can be divided into four distinct epitopes (Watts et al. 1983;Sastry et al,, 1985). A weak, cross-reactive epitope has been localized to the W-terminus (amino acid residues 1-30).…”

The pili of Pseudomonas aeruginosa strains PAK and PAO bind specifically to the carbohydrate sequence βGalNAc(1–4)βGal found in glycosphingolipids asialo‐GM1 and asialo‐GM2

“…The peptides used in these studies have sequences that corresponded to those of residues 128-144 of the PAK (AcPAK(128-144)o«-OH) and PAO (AcPAO(128-144)o^-OH) pilin molecules. The amino acid sequences of the PAK and PAO pilins were derived from Sastry et al (1985). Ac denotes N^-aoetylation of the W-terminus and -OH denotes a peptide with a free a-carboxyi group at the C-terminus.…”

Use of synthetic peptides to confirm that the Pseudomonas aeruginosa PAK pilus adhesin and the Candida albicans fimbrial adhesin possess a homologous receptor‐binding domain

“…Although synonymous, the terms pilus and fimbria are used here to differentiate between the two microorganisms. The P. aeruginosa PAK pilus subunit, pilin, consists of 144 amino acid residues with an intra-chain disulphide loop at its carboxy-terminus (Sastry et al, 1985) that mediates pilus binding to asialo-GM 1 and BEC receptors (Doig et al, 1990;Irvin et al, 1989;Lee et al, 1989a;Paranchych et al, 1986). Although the adhesin is located on the structural subunit of the PAK pilus, it has been shown that the carboxy-terminal region of the PAK pilin is only exposed at the tip of the pilus where receptor binding occurs (Irvin, 1993;Lee et al, 1994).…”

“…Synthetic peptides which corresponded to the different regions of the P. aeruginosa strain PAK pilin sequence (Sastry et al, 1985) were prepared using an Applied Biosystem Inc. model 430A automated peptide synthesizer. The syntheses, purification and characterization of these peptides have been described previously (Wong et al, 1992).…”