- Original Article
- Open Access
Salivary protein profiling in type I diabetes using two-dimensional electrophoresis and mass spectrometry
Clinical Proteomicsvolume 2, pages117–127 (2006)
Owing to its noninvasive collection, saliva is considered as a potent diagnostic fluid. The goal of this study was to investigate the modification of the salivary proteome occurring in type 1 diabetes to highlight potential biomarkers of the pathology. High-resolution two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were combined to perform a largescale analysis. The proteomic comparison of saliva samples from healthy subjects and poorly controlled type 1 diabetes patients revealed a modulation of 23 proteins. Fourteen isoforms of α-amylase, one prolactin inducible protein, three isoforms of salivary acidic protein-1, and three isoforms of salivary cystatins SA-1 were detected as under expressed, whereas two isoforms of serotransferrin were over expressed in the pathological condition. The proteins under expressed were all known to be implicated in the oral anti-inflammatory process, suggesting that the pathology induced a decrease of non-immunological defense of oral cavity. As only particular isoforms of proteins were modulated, type 1 diabetes seemed to differentially affect posttranslational modification.
Kaufman, E. and Lamster, I. B. (2002) The diagnostic applications of saliva-a review. Crit. Rev. Oral. Biol. Med. 13, 197–212.
Lawrence, H. P. (2002) Salivary markers of systemic disease: noninvasive diagnosis of disease and monitoring of general health. J. Can. Dent. Assoc. 68, 170–174.
Streckfus, C. F. and Bigler, L. R. (2002) Saliva as a diagnostic fluid. Oral. Dis. 8, 69–76.
Mata, A. D., Marques, D., Rocha, S., et al. (2004) Effects of diabetes mellitus on salivary secretion and its composition in the human. Mol. Cell. Biochem. 261, 137–142.
Bridges, R. B., Anderson, J. W., Saxe, S. R., Gregory, K., and Bridges, S. R. (1996) Periodontal status of diabetic and non-diabetic men: effects of smoking, glycemic control, and socioeconomic factors. J. Periodontol. 67, 1185–1192.
de Pommereau, V., Dargent-Pare, C., Robert, J. J., and Brion, M. (1992) Periodontal status in insulin-dependent diabetic adolescents. J. Clin. Periodontol. 19, 628–632.
Musumeci, V., Cherubini, P., Zuppi, C., Zappacosta, B., Ghirlanda, G., and Di Salvo, S. (1993) Aminotransferases and lactate dehydrogenase in saliva of diabetic patients. J. Oral. Pathol. Med. 22, 73–76.
Cinquini, I., Calisti, L., Fierabracci, V., et al. (2002) Enzymatic markers of salivary cell injury in saliva of type 1 diabetic children. Clin. Oral. Investig. 6, 21–23.
Todd, A. L., Ng, W. Y., Lee, Y. S., Loke, K. Y., and Thai, A. C. (2002) Evidence of autoantibodies to glutamic acid decarboxylase in oral fluid of type 1 diabetic patients. Diabetes Res. Clin. Pract. 57, 171–177.
Markopoulos, A. K., Belazi, M. A., and Drakoulakos, D. (1997) Glutamic acid decarboxylase autoantibodies in saliva of children with type 1 diabetes. Diabetes Res. Clin. Pract. 38, 169–172.
Yao, Y., Berg, E. A., Costello, C. E., Troxler, R. F., and Oppenheim, F. G. (2003) Identification of protein components in human acquired enamel pellicle and whole saliva using novel proteomics approaches. J. Biol. Chem. 278, 5300–5308.
Ghafouri, B., Tagesson, C., and Lindahl, M. (2003) Mapping of proteins in human saliva using two dimensional gel electrophoresis and peptide mass fingerprinting. Proteomics 3, 1003–1010.
Vitorino R., Lobo, M. J., Duarte, J. A., Ferrer-Correia, A. J., Domingues, P. M., and Amado, F. M. (2004) Identification of human whole saliva protein components using proteomics. Proteomics 4, 1109–1115.
Hardt, M., Thomas, L. R., Dixon, S. E., et al. (2005) Toward defining the human parotid gland salivary proteome and peptidome: identification and characterization using 2D SDS-PAGE, ultrafiltration, HPLC, and mass spectrometry. Biochemistry 44, 2885–2899.
Hu, S., Xie, Y., Ramachandran, P., et al. (2005) Large-scale identification of proteins in human salivary proteome by liquid chromatography/mass spectrometry and two-dimensional gel electrophoresis-mass spectrometry. Proteomics 5, 1714–1728.
Hirtz C., Chevalier, F., Sommerer, N., et al. (2005) Complexity of the human whole saliva proteome. J. Physiol. Biochem. 61, 469–480.
Hirtz, C., Chevalier, F., Centeno, D., et al. (2005) MS characterization of multiple forms of alpha-amylase in human saliva. Proteomics 5, 4597–4607.
Huang, C. M. (2004) Comparative proteomic analysis of human whole saliva. Arch. Oral. Biol. 49, 951–962.
Bradford, M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.
Jenson, O. N., Wilm, M., Srevchenko, A., and Mann, M. (1994) Peptide sequencing of 2-DE gel isolated proteins by nanoelectrospray tandem mass spectrometry. Methods Mol. Biol. 112, 513–530.
Hill, J. (2004) Identifying and managing the complications of diabetes. Nurs. Times 100, 40–44.
Varvarovska, J., Racek, J., Stetina, R., et al. (2004) Aspects of oxidative stress in children with type 1 diabetes mellitus. Biomed. Pharmacother. 58, 539–545.
Bernfeld, P. (1951) Enzymes of starch degradation and synthesis. In Advances in Enzymology Vol. 12, (Nord, F. F., ed.), Interscience, New York, pp. 379–385.
Zakowski, J. J. and Bruns, D. E. (1985) Biochemistry of human alpha amylase isoenzymes. Crit. Rev. Clin. Lab. Sci. 21, 283–322.
Rudney, J. D., Ji, Z., Larson, C. J., Liljemark, W. F., and Hickey, K. L. (1995) Saliva protein binding to layers of oral streptococci in vitro and in vivo. J. Dent. Res. 74, 1280–1288.
Dickinson, D. P. (2002) Cysteine peptidases of mammals: their biological roles and potential effects in the oral cavity and other tissues in health and disease. Crit. Rev. Oral. Biol. Med. 13, 238–275.
Baron, A., DeCarlo, A., and Featherstone, J. (1999) Functional aspects of the human salivary cystatins in the oral environment. Oral. Dis. 5, 234–240.
Schenkels, L. C., Walgreen-Weterings, E., Oomen, L. C., et al. (1997) In vivo binding of the salivary glycoprotein EP-GP (identical to GCDFP-15) to oral and non-oral bacteria detection and identification of EP-GP binding species. Biol. Chem. 378, 83–88.
Clark, J. W., Snell, L., Shiu, R. P., et al. (1999) The potential role for prolactin-inducible protein (PIP) as a marker of human breast cancer micrometastasis. Br. J. Cancer 81, 1002–1008.
Lee, B., Bowden, G. H., and Myal, Y. (2002) Identification of mouse submaxillary gland protein in mouse saliva and its binding to mouse oral bacteria. Arch. Oral. Biol. 47, 327–332.
Morgan, E. H. (1972) The role of transferrin in iron metabolism. Med. J. Aust. 2, 322–325.
Thomas, M. C., MacIsaac, R. J., Tsalamandris, C., and Jerums, G. (2004) Elevated iron indices in patients with diabetes. Diabet. Med. 21, 798–802.
Jones, A. F., Winkles, J. W., Jennings, P. E., Florkowski, C. M., Lunec, J., and Barnett, A. H. (1988) Serum antioxidant activity in diabetes mellitus. Diabetes Res. 7, 89–92.
Vining, R. F., McGinley, R. A., and Symons, R. G. (1983) Hormones in saliva: mode of entry and consequent implications for clinical interpretation. Clin. Chem. 29, 1752–1756.
Lac, G. (2001) Saliva assays in clinical and research biology. Pathol. Biol. 49, 660–667.
Wilmarth, P. A., Riviere, M. A., Rustvold, D. L., Lauten, J. D., Madden, T. E., and David, L. L. (2004) Two-dimensional liquid chromatog raphy study of the human whole saliva proteome. J. Proteome Res. 3, 1017–1023.
Ferguson, D. B. (1987) Current diagnostic uses of saliva. J. Dent. Res. 66, 420–424.
Authors have contributed equally to this work.