Springer Protocols: Abstract: Isolation and Solubilization of Cellular Membrane Proteins from Bacteria

Isolation and Solubilization of Cellular Membrane Proteins from Bacteria

By: Kheir Zuobi-Hasona³, L. Jeannine Brady³

Abstract

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Membrane proteins are rarely identified in two-dimensional electrophoretic (2-DE) proteomics maps. This is because of low abundance, poor solubility, and inherent hydrophobicity. In this study, membrane preparations from the Gram-positive bacterium Streptococcus mutans were isolated from protoplasts and by mechanical grinding. Membrane proteins were extracted using a mixture of trifluroethanol and chloroform, solubilized using highly chaotropic buffer containing ASB-14 and Triton X-100 and subjected to two-dimensional gel electrophoresis.

Affiliation(s): (3) University of Florida, Gainesville, Florida

Book Title: 2D PAGE: Sample Preparation and Fractionation

Series: Methods in Molecular Biology | Volume: 425 | Pub. Date: Jan-25-2008 | Page Range: 287-293 | DOI: 10.1007/978-1-60327-210-0_23

Subject: Protein Science

Key Words: Gram-positive - membrane proteins - solubilization - streptococcus mutans - two-dimensional gel electrophoresis

References

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