Binding of the pili of Pseudomonas aeruginosa to a low-molecular-weight mucin and neutral cystatin of human submandibular-sublingual saliva.
In previous experiments, we have shown that [125I]pili of Pseudomonas aeruginosa exhibited specific binding to a low-molecular-weight mucin (MG2) of human submandibular-sublingual saliva (HSMSL; Reddy MS, Levine MJ, Paranchych W. Crit Rev Oral Biol Med 4:315-323, 1993). In the present study, I have utilized unlabeled pili and immunostaining to identify the receptor molecules in HSMSL. In addition to MG2, pili also bound to neutral cystatin (CsnSN). Binding of unlabeled pili to MG2 and CsnSN could be abolished by treatment of HSMSL with trypsin to hydrolyze the peptide moieties or N-acetylation to neutralize the positive charges of the lysine residues. Reductive methylation of HSMSL, which modifies the lysine residues to methyl lysines while retaining the positive charges, did not affect the binding of pili to either MG2 or CsnSN. Further, pili also exhibited binding to a recombinant MG2 peptide (aa 1-86). Collectively, the data suggested that a protein-to-protein interaction via electrostatic forces mediates the binding of the pili to MG2 and CsnSN. Iodination of pili, which modifies tyrosine-24 and/or -27 residues to O-iodotyrosine(s), abolished its binding to CsnSN but not to MG2. These results suggested that the conformation of pili also plays a role in interaction with CsnSN. Conformational change(s) of pili induced by iodination also made it susceptible to hydrolysis with trypsin.
Reddy
M S
MS
Department of Oral Biology, School of Dental Medicine, State University of New York at Buffalo, Buffalo, NY 14214, USA.
D015891:Cystatins / Q000378:metabolism; D010861:Fimbriae, Bacterial / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D009077:Mucins / Q000378:metabolism; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D012471:Salivary Proteins and Peptides / Q000378:metabolism*; D013361:Sublingual Gland / Q000378:metabolism*; D013363:Submandibular Gland / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology
D015891:Cystatins; C083238:MUC7 protein, human; D009077:Mucins; D012471:Salivary Proteins and Peptides; D014357:Trypsin