Catch-and-release reagents for broadscale quantitative proteomics analyses.

Catch-and-release reagents for broadscale quantitative proteomics analyses. Research Abstract Details 

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Catch-and-release reagents for broadscale quantitative proteomics analyses. Abstract Text:

Carlos A Gartner,Joshua E Elias,Corey E Bakalarski,Steven P Gygi,

The relative quantification of protein expression levels in different cell samples through the utilization of stable isotope dilution has become a standard method in the field of proteomics. We describe here the development of a new reductively cleavable reagent which facilitates the relative quantification of thousands of proteins from only tens of micrograms of starting protein. The ligand features a novel disulfide moiety that links biotin and a thiol-reactive entity. The disulfide is stable to reductive conditions employed during sample labeling but is readily cleaved under mild conditions using tris-(2-carboxyethyl) phosphine (TCEP). This unique chemical property allows for the facile use of immobilized avidin in a manner equivalent to the use of conventional reversible-binding affinity resins. Target peptides are bound to avidin resin, washed rigorously, then cleaved directly from the resin, resulting in simplified sample handling procedures and reduced nonspecific interactions. Here we demonstrate the stability of the linker under two different reducing conditions and show how this "catch-and-release (CAR)" reagent can be used to quantitatively compare protein abundances from two distinct cellular lysates. Starting with only 40 microg protein from each sample, 1840 individual proteins were identified in a single experiment. Using in-house software for automated peak integration, 1620 of these proteins were quantified for differential expression.

Catch-and-release reagents for broadscale quantitative proteomics analyses. Publishing Authors By Initials

CA Gartner,JE Elias,CE Bakalarski,SP Gygi,

For similar biological sciences: biochemistry: proteomics research abstracts see: biological sciences: biochemistry: proteomics research

PUBMED ID PMID:

MEDLINE DATE:

Catch-and-release reagents for broadscale quantitative proteomics analyses. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Journal of proteome research

VOLUME: 6

Page Numbers: 1482-91

Journal Abbreviation:

ISSN: 1535-3893

DAY: 21

MONTH: 02

YEAR: 2007

Catch-and-release reagents for broadscale quantitative proteomics analyses. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 101128775

Catch-and-release reagents for broadscale quantitative proteomics analyses. Keywords Mesh Terms:

KEYWORDS: Proteomics

MESH TERMS: methods

Chemical & Substance for Abstract: Catch-and-release reagents for broadscale quantitative proteomics analyses. Information

Substance Name: tris(2-carboxyethyl)phosphine

Registry Number: 5961-85-3

Grant and Affiliation Information for Catch-and-release reagents for broadscale quantitative proteomics analyses.

AFFILIATION: Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.

Country: United States

AGENCY: United States NHGRI

GRANT: HG003456

ACRONYM: HG

MEDLINETA: J Proteome Res

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