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Detection and first characterization of an uncommon haptoglobin in porcine saliva of pigs with rectal prolapse by using boronic acid sample enrichment

Published online by Cambridge University Press:  10 November 2016

A. M. Gutiérrez ,
I. Miller ,
D. Kolarich ,
K. Hummel ,
K. Nöbauer  and
E. Razzazi-Fazeli
A. M. Gutiérrez*
Affiliation:
Department of Animal Medicine and Surgery, Regional Campus of International Excellence ‘Campus Mare Nostrum’, University of Murcia, 30100 Espinardo, Murcia, Spain
I. Miller
Affiliation:
Department of Biomedical Sciences, University of Veterinary Medicine Vienna, Veterinaerplatz 1, A-1210 Vienna, Austria
D. Kolarich
Affiliation:
Department of Biomolecular Systems, Glycoproteomics Group, Max Planck Institute of Colloids and Interfaces, Arnimallee 22, 14195 Berlin, Germany
K. Hummel
Affiliation:
VetCore Facility for Research, University of Veterinary Medicine Vienna, Veterinaerplatz 1, A-1210 Vienna, Austria
K. Nöbauer
Affiliation:
VetCore Facility for Research, University of Veterinary Medicine Vienna, Veterinaerplatz 1, A-1210 Vienna, Austria
E. Razzazi-Fazeli
Affiliation:
VetCore Facility for Research, University of Veterinary Medicine Vienna, Veterinaerplatz 1, A-1210 Vienna, Austria
*
†E-mail: agmontes@um.es
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Abstract

Salivary glycoprotein profiles, obtained after boronic acid enrichment, were studied for the first time in pigs in order to search for specific overall alterations related to acute inflammatory condition. Five healthy pigs and five pigs suffering from rectal prolapse were used, and the levels of acute phase proteins were measured to determine the degree of inflammation of the animals. The enriched glycoprotein profiles, achieved by two-dimensional gel electrophoresis (2DE) were statistically evaluated and spots that appeared differentially regulated between states were subjected to MS analysis for protein identification. Spots from three unique proteins were identified: carbonic anhydrase VI (CA VI), α-1-antichymotrypsin and haptoglobin (Hp). CA VI appeared as two adjacent horizontal spot trains in the glycoprotein profile of healthy animals in its regular isoelectric points (pI). One spot of α-1-antichymotrypsin was found in saliva from pigs with rectal prolapse in an unusual basic pI, and was considered as a breakdown product. Hp was identified as several spot trains in saliva from pigs with rectal prolapse in an unusual alkaline pI and was consequently further investigated. SDS-PAGE and 2DE of paired serum and saliva samples combined with Western blot analysis showed that the unusual Hp position observed in saliva samples was absent in serum. Furthermore, N-glycans from serum and saliva Hp glycopatterns were evaluated from SDS-PAGE Hp bands and showed that the serum N-glycan distribution in Hp β-chain was comparable in quantity and quality in both groups of animals. In saliva, no Hp β-chain derived N-glycans could unambiguously be identified from this sample set, thus needing further detailed investigations in the future.

Keywords

salivapigglyco-enrichmentproteomicsinflammation
Type
Research Article
Information
animal , Volume 11 , Issue 5 , May 2017 , pp. 845 - 853
Copyright
© The Animal Consortium 2016 

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References

Almeida, A and Kolarich, D 2016. The promise of protein glycosylation for personalised medicine. Biochimica et Biophysica Acta 1860, 1583–1595.Google Scholar
Baldini, C, Giusti, L, Bazzichi, L, Lucacchini, A and Bombardieri, S 2008. Proteomic analysis of the saliva: a clue for understanding primary from secondary Sjögren’s syndrome? Autoimmunity Reviews 7, 185–191.CrossRefGoogle ScholarPubMed
Boonyapranai, K, Tsai, HY, Chen, MC, Sriyam, S, Sinchaikul, S, Phutrakul, S and Chen, ST 2011. Glycoproteomic analysis and molecular modeling of haptoglobin multimers. Electrophoresis 32, 1422–1432.CrossRefGoogle ScholarPubMed
Border, MB, Schwartz, S, Carlson, J, Dibble, CF, Kohltfarber, H, Offenbacher, S, Buse, JB and Bencharit, S 2012. Exploring salivary proteomes in edentulous patients with type 2 diabetes. Molecular Biosystems 8, 1304–1310.Google Scholar
Bradford, MM 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.CrossRefGoogle ScholarPubMed
Fanayan, S, Hincapie, M and Hancock, WS 2012. Using lectins to harvest the plasma/serum glycoproteome. Electrophoresis 33, 1746–1754.Google Scholar
Ferreira, JA, Daniel-da-Silva, AL, Alves, RM, Duarte, D, Vieira, I, Santos, LL, Vitorino, R and Amado, F 2011. Synthesis and optimization of lectin functionalized nanoprobes for the selective recovery of glycoproteins from human body fluids. Analytical Chemistry 83, 7035–7043.CrossRefGoogle ScholarPubMed
Gutiérrez, AM, Martínez-Subiela, S and Cerón, JJ 2009. Evaluation of an immunoassay for determination of haptoglobin concentration in various biological specimens from swine. American Journal of Veterinary Research 70, 691–696.CrossRefGoogle ScholarPubMed
Gutiérrez, AM, Yelamos, J, Pallarés, FJ, Gómez-Laguna, J and Cerón, JJ 2012. Local identification of porcine haptoglobin in salivary gland and diaphragmatic muscle tissues. Histology and Histopathology 27, 187–196.Google ScholarPubMed
Gutiérrez, AM, Escribano, D, Fuentes, M and Cerón, JJ 2013a. Circadian pattern of acute phase proteins in the saliva of growing pigs. Veterinary Journal 196, 167–170.Google Scholar
Gutiérrez, AM, Nöbauer, K, Soler, L, Razzazi-Fazeli, E, Gemeiner, M, Cerón, JJ and Miller, I 2013b. Detection of potential markers for systemic disease in saliva of pigs by proteomics: a pilot study. Veterinary Immunology and Immunopathology 151, 73–82.Google Scholar
Kahlisch, D, Buettner, FF, Naim, HY and Gerlach, GF, FUGATO-consortium IRAS 2009. Glycoprotein analysis of porcine bronchoalveolar lavage fluid reveals potential biomarkers corresponding to resistance to Actinobacillus pleuropneumoniae infection. Veterinary Research 40, 60.Google Scholar
Kolarich, D and Altmann, F 2000. N-glycan analysis by matrix-assisted laser desorption/ionization mass spectrometry of electrophoretically separated nonmammalian proteins: application to peanut allergen Ara h 1 and olive pollen allergen Ole e 1. Analytical Biochemistry 285, 64–75.CrossRefGoogle ScholarPubMed
Kolarich, D, Weber, A, Turecek, PL, Schwarz, HP and Altmann, F 2006. Comprehensive glyco-proteomic analysis of human alpha1-antitrypsin and its charge isoforms. Proteomics 6, 3369–3380.CrossRefGoogle ScholarPubMed
Kushner, I and Mackiewicz, A 1993. The acute phase response: an overview. In Acute phase protein, molecular biology, biochemistry and clinical applications (ed. A Mackiewicz, I Kushner and H Baumawn), pp. 3–19. CDC Press, London.Google Scholar
Loke, I, Kolarich, D, Packer, NH and Thaysen-Andersen, M 2016. Emerging roles of protein mannosylation in inflammation and infection. Molecular Aspects of Medicine 51, 31–55.Google Scholar
Marco-Ramell, A, Miller, I, Nöbauer, K, Möginger, U, Segalés, J, Razzazi-Fazeli, E, Kolarich, D and Bassols, A 2014. Proteomics on porcine haptoglobin and IgG/IgA show protein species distribution and glycosylation pattern to remain similar in PCV2-SD infection. Journal of Proteomics 101, 205–216.CrossRefGoogle ScholarPubMed
Miller, I, Crawford, J and Gianazza, E 2006. Protein stains for proteomic applications: which, when, why? Proteomics 6, 5385–5408.CrossRefGoogle Scholar
Miller, I, Wait, R, Sipos, W and Gemeiner, M 2009. A proteomic reference map for pig serum proteins as a prerequisite for diagnostic applications. Research in Veterinary Sciences 86, 362–367.CrossRefGoogle ScholarPubMed
Moremen, KW, Tiemeyer, M and Nairn, AV 2012. Vertebrate protein glycosylation: diversity, synthesis and function. Nature Reviews. Molecular Cell Biology 13, 448–462.Google Scholar
Nishita, T, Yatsu, J, Watanabe, K, Ochiai, H, Ichihara, N, Orito, K and Arishima, K 2014. Urinary carbonic anhydrase VI as a biomarker for kidney disease in pigs. Veterinary Journal 202, 378–380.CrossRefGoogle ScholarPubMed
Novotny, MV, Alley, WR Jr and Mann, BF 2012. Analytical glycobiology at high sensitivity: current approaches and directions. Glycoconjugate Journal 30, 89–117.CrossRefGoogle ScholarPubMed
Ongay, S, Boichenko, A, Govorukhina, N and Bischoff, R 2012. Glycopeptide enrichment and separation for protein glycosylation analysis. Journal of Separation Science 35, 2341–2372.Google Scholar
Ramachandran, P, Boontheung, P, Pang, E, Yan, W, Wong, DT and Loo, JA 2008. Comparison of N-linked glycoproteins in human whole saliva, parotid, submandibular, and sublingual glandular secretions identified using hydrazide chemistry and mass spectrometry. Clinical Proteomics 4, 80–104.Google Scholar
Reiding, KR, Blank, D, Kuijper, DM, Deelder, AM and Wuhrer, M 2014. High-throughput profiling of protein N-glycosylation by MALDI-TOF-MS employing linkage-specific sialic acid esterification. Analytical Chemistry 86, 5784–5793.Google Scholar
Shevchenko, A, Wilm, M, Vorm, O and Mann, M 1996. Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Analytical Chemistry 68, 850–858.CrossRefGoogle ScholarPubMed
Sok, J, Wang, XZ, Batchvarova, N, Kuroda, M, Harding, H and Ron, D 1999. CHOP-dependent stress-inducible expression of a novel form of carbonic anhydrase VI. Molecular and Cellular Biology 19, 495–504.CrossRefGoogle ScholarPubMed
Sondej, M, Denny, PA, Xie, Y, Ramachandran, P, Si, Y, Takashima, J, Shi, W, Wong, DT, Loo, JA and Denny, PC 2009. Glycoprofiling of the Human Salivary Proteome. Clinical Proteomics 5, 52–68.Google Scholar
Song, B, Zhang, L, Liu, XJ, Ding, C, Wu, LL, Gan, YH and Yu, GY 2012. Proteomic analysis of secretion from human transplanted submandibular gland replacing lacrimal gland with severe keratoconjunctivitis sicca. Biochimica et Biophysica Acta 1824, 550–560.CrossRefGoogle ScholarPubMed
Stratil, A, Cízová-Schröffelová, D, Gábrisová, E, Pavlík, M, Coppieters, W, Peelman, L, Van de Weghe, A and Bouquet, Y 1995. Pig plasma alpha-protease inhibitors PI2, PI3 and PI4 are members of the antichymotrypsin family. Comparative Biochemistry and Physiology B 111, 53–60.CrossRefGoogle ScholarPubMed
Thaysen-Andersen, M, Thøgersen, IB, Lademann, U, Offenberg, H, Giessing, AM, Enghild, JJ, Nielsen, HJ, Brünner, N and Højrup, P 2008. Investigating the biomarker potential of glycoproteins using comparative glycoprofiling – application to tissue inhibitor of metalloproteinases-1. Biochimica et Biophysica Acta 1784, 455–463.CrossRefGoogle ScholarPubMed
Theodoratou, E, Campbell, H, Ventham, NT, Kolarich, D, Pučić-Baković, M, Zoldoš, V, Fernandes, D, Pemberton, IK, Rudan, I, Kennedy, NA, Wuhrer, M, Nimmo, E, Annese, V, McGovern, DP, Satsangi, J and Lauc, G 2014. The role of glycosylation in IBD. Nature Reviews. Gastroenterology & Hepatology 11, 588–600.Google Scholar
Váradi, C, Mittermayr, S, Szekrényes, Á, Kádas, J, Takacs, L, Kurucz, I and Guttman, A 2013. Analysis of haptoglobin N-glycome alterations in inflammatory and malignant lung diseases by capillary electrophoresis. Electrophoresis 34, 2287–12294.CrossRefGoogle ScholarPubMed
Wang, X, Xia, N and Liu, L 2013. Boronic acid-based approach for separation and immobilization of glycoproteins and its application in sensing. International Journal of Molecular Science 14, 20890–20912.Google Scholar
Xu, Y, Bailey, UM, Punyadeera, C and Schulz, BL 2014. Identification of salivary N-glycoproteins and measurement of glycosylation site occupancy by boronate glycoprotein enrichment and liquid chromatography/electrospray ionization tandem mass spectrometry. Rapid Communications in Mass Spectrometry 28, 471–482.CrossRefGoogle ScholarPubMed