Quantitative profiling of differentiation-induced microsomal proteins using isotope-coded affinity tags and mass spectrometry.

Abstract:

:An approach to the systematic identification and quantification of the proteins contained in the microsomal fraction of cells is described. It consists of three steps: (1) preparation of microsomal fractions from cells or tissues representing different states; (2) covalent tagging of the proteins with isotope-coded affinity tag (ICAT) reagents followed by proteolysis of the combined labeled protein samples; and (3) isolation, identification, and quantification of the tagged peptides by multidimensional chromatography, automated tandem mass spectrometry, and computational analysis of the obtained data. The method was used to identify and determine the ratios of abundance of each of 491 proteins contained in the microsomal fractions of naïve and in vitro- differentiated human myeloid leukemia (HL-60) cells. The method and the new software tools to support it are well suited to the large-scale, quantitative analysis of membrane proteins and other classes of proteins that have been refractory to standard proteomics technology.

journal_name

Nat Biotechnol

journal_title

Nature biotechnology

authors

Han DK,Eng J,Zhou H,Aebersold R

doi

10.1038/nbt1001-946

subject

Has Abstract

pub_date

2001-10-01 00:00:00

pages

946-51

issue

10

eissn

1087-0156

issn

1546-1696

pii

nbt1001-946

journal_volume

19

pub_type

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