In order to know the characteristic roles of salivary protein complex (SPC) the gel-filtration chromatography was performed using the unstimulated and the stimulated whole saliva separately. The first and second dominant SPC peaks were fractionated and analyzed by immunoprecipitation HPLC (IP-HPLC) using antibodies against the essential salivary proteins including α-amylase, mucin-1, proline rich proteins (PRPs), histatin, cystatin, LL-37, lysozyme, lactoferrin, -defensin-1, -2, -3, IgA, transglutaminase 4 (TGase 4), mucocidin, α1-antitrypsin, cathepsin G. In the gel-filtration chromatography the stimulated whole saliva showed much reduced amount of SPCs than the unstimulated whole saliva, but the proportional patterns of both whole saliva were almost similar each other. Through IP-HPLC analysis both of the first and second dominant SPCs were variably positive for the essential salivary proteins, however, α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G were predominant in the first dominant SPC, while cystatin, lactoferrin, β-defensin-1, -2,-3, IgA, mucocidin, TGase 4, and α1-antitrypsin were predominant in the second dominant SPC. And more, the α1-antitrypsin and cathepsin G which were mostly derived from gingival crevicular fluid were also consistently found in the SPCs. These data may suggest that the first dominant SPC, rich in α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G, may play a role in food digestion, protein degradation, and mucosa lubrication, while the second dominant SPC, rich in cystatin, lactoferrin, β-defensin-1, -2, -3, mucocidin, IgA, TGase 4, and α1-antitrypsin, may play a role in the mucosa protection and antimicrobial defense.

Human saliva contains a large number of proteins and peptides whose composition may alter as a consequence 。f disease. To date. however‘ the proteins and peptides that routinely populate thi s ora l fluid a re largely unknown To provide a catalogue of saliva proteins, we have surveyed the unstimulated human whole saliva by us ing shotgun proteornics. For the shotgun approach, whole saliva proteins were digested into peptides with ChemDigestD ‘ and the resul ting peptide fragments were separated by RP-HPLC, followed by each fraction was t ryptic digestion. ChemDigestD-Trypsin digested peptides were analyzed by tandem mass spectrometry(MS/MS) us ing a nano-LC eq 버 pped quadrupole-time of f1ight mass spectrometer, and the obtained spectra were searched against human protein seq uence data base using MASCOT. Shotgun proteomics allowed a total of 291 human pr。 teins to be confidently assigned . The largest group(17 .2%) of the identified proteins sorted into functional catego ries was included in t he signal t ransduction funct ion except for the hypothetical 0 1' unknown function. This work provides a valuable s ta rting point for the analysis of human salivary proteins and their biological functions and candidates from human whole sali va that may prove to be of diagnostic and therapeutic significance

Human saliva conta ins a la rge number of proteins and peptides whose composition may alter as a conseq uence of disease. '1'0 date‘ however. the proteins and peptides that routinely populate t his oral fluid are largely unknown, '1'0 provid e a ca ta logue 이， sali va protei ns. we have surveyed the unstimulated human whole saliva by using shotgun proteomics. F'or the shotgun a pproach‘ whole sali va proteins were digested into peptides with ChemDigestD and the res ulting pe ptide fragments were sepa rated by RP- HPLC, followed by each fraction was tryptic di gestion ChemDiges tD- Trypsin diges ted pe pt ides were analyzed by tandem mass spectrometry (MS!MS) using a nano-LC equi pped quadru po le-time of fli ght rnass spect rometer, and the obtained spectra were searched against human protein sequence da tabase us ing MASCOT Shotgun proteomics a llowed a total of 291 human proteins to be confidently assigned. The largest gro u p (17 , 2%) of the identifi ed proteins sorted into functional categories was included in the s ignal transducti on function except for the hypothet ical or unknown functio n, This work provides a valuable starting point for the ana lys is of human sa l i va ry protei ns a nd theil‘ biological functions and candidates from human whole saliva that may prove to be of diagn ost ic and t herapeutic s ignif‘ Ica nce