Large-scale identification of membrane proteins based on analysis of trypsin-protected transmembrane segments

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F16%3A00467847" target="_blank" >RIV/61388971:_____/16:00467847 - isvavai.cz</a>

Alternative codes found

RIV/00216208:11310/16:10326338 RIV/00216208:11110/16:10326338

Result on the web

<a href="http://dx.doi.org/10.1016/j.jprot.2016.03.016" target="_blank" >http://dx.doi.org/10.1016/j.jprot.2016.03.016</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.jprot.2016.03.016" target="_blank" >10.1016/j.jprot.2016.03.016</a>

Result language

angličtina

Original language name

Large-scale identification of membrane proteins based on analysis of trypsin-protected transmembrane segments

Original language description

ntegral membrane proteins are generally under-represented in routine proteomic analyses, mostly because of their relatively low abundance, hydrophobicity and lack of trypsin-cleavage sites. To increase the coverage of membrane proteomes, various strategies have been developed, targeting mostly the extra-membrane segments of membrane proteins. We focused our attention to the rather overlooked hydrophobic transmembrane segments. Such peptides can be isolated after carbonate stripping and protease "shaving" of membranes isolated by simple centrifugation procedure. The treated membranes with embedded hydrophobic peptides can then be solubilized in organic solvents, re-digested with CNBr, delipidated and subjected to LC-MS/MS analysis. We modified the original "hppK" method, and applied it for the analysis of human lymphoma cells. We identified 1224 proteins of which two-thirds were IMPs with 1-16 transmembrane segments. This method allowed us to identify 13 "missing proteins" - proteins with no previous evidence on protein level. nBiological significance: Integral membrane proteins execute numerous essential functions and represent substantial part of eukaryotic proteomes. Our knowledge of their function and expression is, however, limited. Novel approaches extending our knowledge of membrane proteome are therefore highly desired. As we demonstrate here, a non-conventional method which targets rather overlooked hydrophobic transmembrane segments of integral membrane proteins has wide potential to provide the missing information on the membrane proteome. We show that it can deliver identification and potentially also quantification of hundreds of integral membrane proteins including the so called "missing proteins".

Czech name

—

Czech description

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Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CE - Biochemistry

OECD FORD branch

—

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2016

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Proteomics

ISSN

1874-3919

e-ISSN

—

Volume of the periodical

146

Issue of the periodical within the volume

SI

Country of publishing house

GB - UNITED KINGDOM

Number of pages

8

Pages from-to

15-22

UT code for WoS article

000387522800004

EID of the result in the Scopus database

2-s2.0-84961901410