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


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2001-10

AUTHORS

David K. Han, Jimmy Eng, Huilin Zhou, Ruedi Aebersold

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. More... »

PAGES

946-951

Journal

TITLE

Nature Biotechnology

ISSUE

10

VOLUME

19

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nbt1001-946

    DOI

    http://dx.doi.org/10.1038/nbt1001-946

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1029399357

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/11581660


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