A simplified method for peptide de novo sequencing using O-18
labeling

J 2014

A simplified method for peptide de novo sequencing using O-18 labeling

VORÁČ, Aleš, Ondrej ŠEDO, Jan HAVLIŠ a Zbyněk ZDRÁHAL

Základní údaje

Originální název

A simplified method for peptide de novo sequencing using O-18 labeling

Autoři

VORÁČ, Aleš (203 Česká republika, domácí), Ondrej ŠEDO (203 Česká republika, domácí), Jan HAVLIŠ (203 Česká republika, domácí) a Zbyněk ZDRÁHAL (203 Česká republika, garant, domácí)

Vydání

European Journal of Mass Spectrometry, CHICHESTER (ENGLAND), IM PUBLICATIONS, 2014, 1469-0667

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 1.000

Kód RIV

RIV/00216224:14740/14:00073785

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1255/ejms.1277

UT WoS

000337268400006

Klíčová slova anglicky

peptide de novo sequencing; mass spectrometry; isotopic labeling; O-18 incorporation

Štítky

kontrola MP, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 11. 3. 2015 11:30, Martina Prášilová

Anotace

V originále

Incorporation of an O-18 atom into a peptide C-terminus by proteolytic cleavage in the presence of (H2O)-O-18 is one of the most effective ways of enhancing tandem mass spectrometry (MS/MS)-based de novo sequencing. Incorporation is usually accomplished by procedures including vacuum-assisted drying of tryptic peptides extracted from gels, their subsequent reconstitution in a (H2O)-O-16/(H2O)-O-18 mixture and re-treatment with trypsin. In the present work, we propose a simplified procedure for O-18 incorporation into tryptic peptides by adding (H2O)-O-18 and trypsin to the original digest solution. In comparison to published methods, the proposed protocol for peptide de novo sequencing brings significant advantages in analysis and workflow with no deterioration in method performance. We show that labeling by this simplified method leads to a highlighting of the y-ion fragment series in the peptide matrix-assisted laser desorption/ionization (MALDI)-MS/MS data, which facilitates MS/MS data interpretation. We also prove that eliminating acid extraction of peptides from gels does not result in a decrease in sequence coverage or a qualitative loss of particular peptides detectable by MALDI-MS. The method was examined by MALDI-MS/MS on bovine serum albumin and recombinant histidine kinase CKI1 from Arabidopsis thaliana, and was verified by de novo sequencing of tryptic peptides originating from Apodemus sylvaticus salivary proteins.

Návaznosti

ED1.1.00/02.0068, projekt VaV

Název: CEITEC - central european institute of technology

GBP206/12/G151, projekt VaV

Název: Centrum nových přístupů k bioanalýze a molekulární diagnostice

Citovat

VORÁČ, Aleš, Ondrej ŠEDO, Jan HAVLIŠ a Zbyněk ZDRÁHAL. A simplified method for peptide de novo sequencing using O-18 labeling. European Journal of Mass Spectrometry. CHICHESTER (ENGLAND): IM PUBLICATIONS, 2014, roč. 20, č. 3, s. 255-260. ISSN 1469-0667. Dostupné z: https://dx.doi.org/10.1255/ejms.1277.

@article{1194851,
   author = {Voráč, Aleš and Šedo, Ondrej and Havliš, Jan and Zdráhal, Zbyněk},
   article_location = {CHICHESTER (ENGLAND)},
   article_number = {3},
   doi = {http://dx.doi.org/10.1255/ejms.1277},
   keywords = {peptide de novo sequencing; mass spectrometry; isotopic labeling; O-18 incorporation},
   language = {eng},
   issn = {1469-0667},
   journal = {European Journal of Mass Spectrometry},
   title = {A simplified method for peptide de novo sequencing using O-18 labeling},
   url = {https://www.impublications.com/content/abstract?code=E20_0255},
   volume = {20},
   year = {2014}
}

TY  - JOUR
ID  - 1194851
AU  - Voráč, Aleš - Šedo, Ondrej - Havliš, Jan - Zdráhal, Zbyněk
PY  - 2014
TI  - A simplified method for peptide de novo sequencing using O-18 labeling
JF  - European Journal of Mass Spectrometry
VL  - 20
IS  - 3
SP  - 255-260
EP  - 255-260
PB  - IM PUBLICATIONS
SN  - 14690667
KW  - peptide de novo sequencing
KW  - mass spectrometry
KW  - isotopic labeling
KW  - O-18 incorporation
UR  - https://www.impublications.com/content/abstract?code=E20_0255
L2  - https://www.impublications.com/content/abstract?code=E20_0255
N2  - Incorporation of an O-18 atom into a peptide C-terminus by proteolytic cleavage in the presence of (H2O)-O-18 is one of the most effective ways of enhancing tandem mass spectrometry (MS/MS)-based de novo sequencing. Incorporation is usually accomplished by procedures including vacuum-assisted drying of tryptic peptides extracted from gels, their subsequent reconstitution in a (H2O)-O-16/(H2O)-O-18 mixture and re-treatment with trypsin. In the present work, we propose a simplified procedure for O-18 incorporation into tryptic peptides by adding (H2O)-O-18 and trypsin to the original digest solution. In comparison to published methods, the proposed protocol for peptide de novo sequencing brings significant advantages in analysis and workflow with no deterioration in method performance. We show that labeling by this simplified method leads to a highlighting of the y-ion fragment series in the peptide matrix-assisted laser desorption/ionization (MALDI)-MS/MS data, which facilitates MS/MS data interpretation. We also prove that eliminating acid extraction of peptides from gels does not result in a decrease in sequence coverage or a qualitative loss of particular peptides detectable by MALDI-MS. The method was examined by MALDI-MS/MS on bovine serum albumin and recombinant histidine kinase CKI1 from Arabidopsis thaliana, and was verified by de novo sequencing of tryptic peptides originating from Apodemus sylvaticus salivary proteins.
ER  -

VORÁČ, Aleš, Ondrej ŠEDO, Jan HAVLIŠ a Zbyněk ZDRÁHAL. A simplified method for peptide de novo sequencing using O-18 labeling. \textit{European Journal of Mass Spectrometry}. CHICHESTER (ENGLAND): IM PUBLICATIONS, 2014, roč.~20, č.~3, s.~255-260. ISSN~1469-0667. Dostupné z: https://dx.doi.org/10.1255/ejms.1277.

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