Technical report: In-gel sample preparation prior to proteomic analysis of bovine faeces increases protein identifications by removal of high molecular weight glycoproteins

Huang, Y., Burchmore, R. , Jonsson, N. N. , Johnson, P. C.D. and Eckersall, P. D. (2022) Technical report: In-gel sample preparation prior to proteomic analysis of bovine faeces increases protein identifications by removal of high molecular weight glycoproteins. Journal of Proteomics, 261, 104573. (doi: 10.1016/j.jprot.2022.104573) (PMID:35351658)

Abstract

Bovine faecal composition is complex and a knowledge gap exists in the understanding of the bovine faecal proteome. In the present study, in-gel sample preparation (IGSP) of faecal samples prior to proteomics showed an increase in the number of proteins identified in faecal samples compared to those processed by filter-aided sample preparation (FASP). The optimised sample preparation method removed high molecular weight glycoproteins as part of the clean-up process of the faecal samples, and in combination with in-gel digestion before liquid chromatography with tandem mass spectrometry (LC-MS/MS) The use of IGSP led to enhanced protein identification with increases in the number of peptides identified and in the percent coverage of proteins in the bovine faecal samples. Significance: Characterization of faecal proteins has the potential to increase our understanding of host responses to changes such as diet, disease and drug-treatment. In-gel sample preparation prior to proteomics can be used to remove high molecular weight glycoproteins and reduce protein/peptide loss in FASP. This method of sample preparation will have application not only in the investigation of bovine faecal extracts but also in studies where large molecules such as glycoproteins or oligosaccharides could have detrimental influences on sample preparation involving ultrafiltration.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Jonsson, Professor Nicholas and Burchmore, Dr Richard and Eckersall, Professor David and Johnson, Dr Paul and Huang, Yixin
• Authors: Huang, Y., Burchmore, R., Jonsson, N. N., Johnson, P. C.D., and Eckersall, P. D.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity; College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
• Journal Name: Journal of Proteomics
• Publisher: Elsevier
• ISSN: 1874-3919
• ISSN (Online): 1876-7737
• Published Online: 26 March 2022
• Copyright Holders: Copyright © 2022 Elsevier B.V.
• First Published: First published in Journal of Proteomics 261: 104573
• Publisher Policy: Reproduced in accordance with the publisher copyright policy