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 Biotechnoljournal_title
Nature biotechnologyauthors
Han DK,Eng J,Zhou H,Aebersold Rdoi
10.1038/nbt1001-946subject
Has Abstractpub_date
2001-10-01 00:00:00pages
946-51issue
10eissn
1087-0156issn
1546-1696pii
nbt1001-946journal_volume
19pub_type
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