Sökning: onr:"swepub:oai:lup.lub.lu.se:bc76bb03-e834-4ba6-abdb-afe1a0776e81" >
Enzymatic generatio...
-
Ebhardt, H AlexanderETH Zürich
(författare)
Enzymatic generation of peptides flanked by basic amino acids to obtain MS/MS spectra with 2× sequence coverage
- Artikel/kapitelEngelska2014
Förlag, utgivningsår, omfång ...
-
2014-11-07
-
Wiley,2014
-
9 s.
Nummerbeteckningar
-
LIBRIS-ID:oai:lup.lub.lu.se:bc76bb03-e834-4ba6-abdb-afe1a0776e81
-
https://lup.lub.lu.se/record/bc76bb03-e834-4ba6-abdb-afe1a0776e81URI
-
https://doi.org/10.1002/rcm.7069DOI
Kompletterande språkuppgifter
-
Språk:engelska
-
Sammanfattning på:engelska
Ingår i deldatabas
Klassifikation
-
Ämneskategori:art swepub-publicationtype
-
Ämneskategori:ref swepub-contenttype
Anmärkningar
-
RATIONALE: Tandem mass (MS/MS) spectra generated by collision-induced dissociation (CID) typically lack redundant peptide sequence information in the form of e.g. b- and y-ion series due to frequent use of sequence-specific endopeptidases cleaving C- or N-terminal to Arg or Lys residues.METHODS: Here we introduce arginyl-tRNA protein transferase (ATE, EC 2.3.2.8) for proteomics. ATE recognizes acidic amino acids or oxidized Cys at the N-terminus of a substrate peptide and conjugates an arginine from an aminoacylated tRNA(Arg) onto the N-terminus of the substrate peptide. This enzymatic reaction is carried out under physiological conditions and, in combination with Lys-C/Asp-N double digest, results in arginylated peptides with basic amino acids on both termini.RESULTS: We demonstrate that in vitro arginylation of peptides using yeast arginyl tRNA protein transferase 1 (yATE1) is a robust enzymatic reaction, specific to only modifying N-terminal acidic amino acids. Precursors originating from arginylated peptides generally have an increased protonation state compared with their non-arginylated forms. Furthermore, the product ion spectra of arginylated peptides show near complete 2× fragment ladders within the same MS/MS spectrum using commonly available electrospray ionization peptide fragmentation modes. Unexpectedly, arginylated peptides generate complete y- and c-ion series using electron transfer dissociation (ETD) despite having an internal proline residue.CONCLUSIONS: We introduce a rapid enzymatic method to generate peptides flanked on either terminus by basic amino acids, resulting in a rich, redundant MS/MS fragment pattern.
Ämnesord och genrebeteckningar
-
Amino Acids, Basic
-
Aminoacyltransferases
-
Peptides
-
Sequence Analysis, Protein
-
Tandem Mass Spectrometry
Biuppslag (personer, institutioner, konferenser, titlar ...)
-
Nan, JieLund University,Lunds universitet,MAX IV-laboratoriet,MAX IV Laboratory(Swepub:lu)maxl-jna
(författare)
-
Chaulk, Steven GUniversity of Alberta
(författare)
-
Fahlman, Richard PUniversity of Alberta
(författare)
-
Aebersold, RuediETH Zürich
(författare)
-
ETH ZürichMAX IV-laboratoriet
(creator_code:org_t)
Sammanhörande titlar
-
Ingår i:Rapid Communications in Mass Spectrometry: Wiley28:24, s. 43-27351097-02310951-4198
Internetlänk
Hitta via bibliotek
Till lärosätets databas