کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1226203 968285 2009 12 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A proteomics approach to study in vivo protein Nα-modifications
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
A proteomics approach to study in vivo protein Nα-modifications
چکیده انگلیسی

In this article we present a simple method to enrich peptides containing in vivo Nα-modified protein N-termini. We demonstrate that CNBr-activated Sepharose, a commercial amine reactive matrix, can selectively couple peptides via the α-NH2 group under mild conditions. Following digestion by trypsin, a simple incubation step with the CNBr-activated Sepharose by which the free α-NH2 containing peptides are coupled with matrix through a covalent bond, allows the separation of Nα-modified peptides from massive free α-NH2 containing peptides. The removal of contaminant peptides with artificial Nα-modifications, like cyclization of N-terminal S-carbamoylmethylcysteine and glutamine, are also discussed. Application of this method to tryptic digests of HeLa cell proteins resulted by a single LC-MS/MS analysis in the identification of 588 in vivo Nα-modified peptides, of which 507 contain IPI (International Protein Index) annotated protein N-termini and 81 contain IPI unannotated protein N-termini. Most of the identified modifications are acetylations with only a few formylations and propionylations present. Furthermore, Lys-N digestion was also applied and resulted in the identification of 394 in vivo Nα-modified peptides, of which 371 contain IPI annotated protein N-termini and 23 contain IPI unannotated protein N-termini. Combination of the two datasets leads to the identification of 675 Nα-modified IPI annotated protein N-termini and 88 Nα-modified IPI unannotated protein N-termini. Our results suggest that N-terminal acetyltransferases (NATs) may function as N-terminal formyltransferases (NFTs) and N-terminal propionyltransferases (NPTs) in vivo.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Proteomics - Volume 73, Issue 2, 1 December 2009, Pages 240–251
نویسندگان
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