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Protein Extraction and 2-DE of Water- And Lipid-Soluble Proteins From Bovine Pericardium, a Low-Cellularity Tissue

Protein Extraction and 2-DE of Water- And Lipid-Soluble Proteins From Bovine Pericardium, a Low-Cellularity Tissue

Leigh G Griffiths, Leila Choe, Kelvin H Lee, Kenneth F Reardon, E Christopher Orton

Electrophoresis. 2008 Nov;29(22):4508-15.

PMID: 18985661

Abstract:

Bovine pericardium (BP) is an important biomaterial used in the production of glutaraldehyde-fixed heart valves and tissue-engineering applications. The ability to perform proteomic analysis on BP is useful for a range of studies, including investigation of immune rejection after implantation. However, proteomic analysis of fibrous tissues such as BP is challenging due to their relative low-cellularity and abundance of extracellular matrix. A variety of methods for tissue treatment, protein extraction, and fractionation were investigated with the aim of producing high-quality 2-DE gels for both water- and lipid-soluble BP proteins. Extraction of water-soluble proteins with 3-(benzyldimethylammonio)-propanesulfonate followed by n-dodecyl beta-D-maltoside extraction and ethanol precipitation for lipid-soluble proteins provided the best combination of yield, spot number, and resolution on 2-DE gels (Protocol E2). ESI-quadrupole/ion trap or MALDI-TOF/TOF MS protein identifications were performed to confirm bovine origin and appropriate subcellular prefractionation of resolved proteins. Twenty-five unique, predominantly cytoplasmic bovine proteins were identified from the water-soluble fraction. Thirty-two unique, predominantly membrane bovine proteins were identified from the lipid-soluble fraction. These results demonstrated that the final protocol produced high-quality proteomic data from this important tissue for both cytoplasmic and membrane proteins.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP81239454	3-(Benzyldimethylammonio)propanesulfonate		81239-45-4	Price

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