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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: [email protected]
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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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