Work

Subcellular Fractionation and Proteomics of Nuclear Envelopes

Year: 2008

Type: book-chapter

Abstract: Because of its many connections to other cell systems, the nuclear envelope (NE)is essentially impossible to purify to homogeneity. To circumvent these problems, we developed a subtractive proteomics approach in which the fraction of interest and a fraction known to contaminate the fraction of interest are separately analyzed, and proteins identified in both fractions are subtracted from the data set. This requires that the contaminating fraction can be purified to homogeneity. In this case, microsomal membranes (MMs) are used to represent endoplasmic reticulum contamination, allowing the identification of transmembrane proteins specific to the NE. To circumvent problems commonly associated with analyzing membrane proteins, the multidimensional protein identification technology (MudPIT) proteomics methodology is employed.

Source: Methods in molecular biology

DOI: 10.1007/978-1-59745-028-7_8

Authors: Laurence Florens, Nadia Korfali, Eric C. Schirmer

Institutions: Stowers Institute for Medical Research, Wellcome Centre for Cell Biology, University of Edinburgh

Language: English

Topic: Advanced Proteomics Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: green

Funder: Wellcome Trust

Award: The role of the nuclear envelope in genome architecture and cellular differentiation.