PLUNC in human nasal lavage fluid: multiple isoforms that bind to lipopolysaccharide.
Here, we demonstrate the presence of multiple isoforms of palate lung nasal epithelial clone (PLUNC) in human nasal lavage fluid (NLF). Eight isoforms were separated by two-dimensional gel electrophoresis (2-DE), and peptide mapping of the proteins was performed using MALDI-TOF MS (matrix assisted laser desorption/ionization time of flight mass spectrometry) of tryptic and asparginase cleavages. The identification was verified by amino acid sequencing after analysis of collision-induced dissociation (CID) fragmentation spectra with nanoelectrospray MS/MS. One isoform showed an electrophoretic mobility shift after N-glycosidase treatment, indicating that at least one of the PLUNC isoforms is glycosylated. We also demonstrate that PLUNC in NLF binds to lipopolysaccharide (LPS) in vitro; indeed, out of all proteins present in NLF only the PLUNC isoforms were found to adsorb to an LPS-coated surface. These results show that PLUNC is expressed as multiple LPS-binding isoforms in human NLF. The possibility that PLUNC may play a role in the innate immune response of the upper airways is inferred.
Ghafouri
Bijar
B
Division of Occupational and Environmental Medicine, Department of Molecular and Clinical Medicine, Faculty of Health Sciences, Linköping University, S-581 85 Linköping, Sweden.
Kihlström
Erik
E
Tagesson
Christer
C
Lindahl
Mats
M
D015180:Electrophoresis, Gel, Two-Dimensional; D024202:Electrophoretic Mobility Shift Assay; D006023:Glycoproteins / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D016350:Leucine Zippers; D008070:Lipopolysaccharides / Q000378:metabolism*; D017817:Nasal Lavage Fluid / Q000737:chemistry*; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000032:analysis* / Q000378:metabolism; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism; D020545:Respiratory Mucosa / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
C485599:BPIFA1 protein, human; D006023:Glycoproteins; D008070:Lipopolysaccharides; D010750:Phosphoproteins; D020033:Protein Isoforms