Protocol for absolute quantification of proteins in Gram-negative bacteria based on QconCAT-based labeled peptides

Mass-spectrometry-based absolute protein quantification uses labeled quantification concatamer (QconCAT) as internal standards (ISs). To calculate the amount of protein(s), the ion intensity ratio between the analyte and its cognate IS is compared in each biological sample. The present protocol desc...

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Detalles Bibliográficos
Autores principales: Gurdo, Nicolás, Taylor Parkins, Shannara Kayleigh, Fricano, Martina, Wulff, Tune, Nielsen, Lars Keld, Nikel, Pablo Iván
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9881405/
https://www.ncbi.nlm.nih.gov/pubmed/36853682
http://dx.doi.org/10.1016/j.xpro.2023.102060
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author Gurdo, Nicolás
Taylor Parkins, Shannara Kayleigh
Fricano, Martina
Wulff, Tune
Nielsen, Lars Keld
Nikel, Pablo Iván
author_facet Gurdo, Nicolás
Taylor Parkins, Shannara Kayleigh
Fricano, Martina
Wulff, Tune
Nielsen, Lars Keld
Nikel, Pablo Iván
author_sort Gurdo, Nicolás
collection PubMed
description Mass-spectrometry-based absolute protein quantification uses labeled quantification concatamer (QconCAT) as internal standards (ISs). To calculate the amount of protein(s), the ion intensity ratio between the analyte and its cognate IS is compared in each biological sample. The present protocol describes a systematic workflow to design, produce, and purify QconCATs and to quantify soluble proteins in Pseudomonas putida KT2440. Our methodology enables the quantification of detectable peptide and serves as a versatile platform to produce ISs for different biological systems.
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spelling pubmed-98814052023-01-28 Protocol for absolute quantification of proteins in Gram-negative bacteria based on QconCAT-based labeled peptides Gurdo, Nicolás Taylor Parkins, Shannara Kayleigh Fricano, Martina Wulff, Tune Nielsen, Lars Keld Nikel, Pablo Iván STAR Protoc Protocol Mass-spectrometry-based absolute protein quantification uses labeled quantification concatamer (QconCAT) as internal standards (ISs). To calculate the amount of protein(s), the ion intensity ratio between the analyte and its cognate IS is compared in each biological sample. The present protocol describes a systematic workflow to design, produce, and purify QconCATs and to quantify soluble proteins in Pseudomonas putida KT2440. Our methodology enables the quantification of detectable peptide and serves as a versatile platform to produce ISs for different biological systems. Elsevier 2023-01-24 /pmc/articles/PMC9881405/ /pubmed/36853682 http://dx.doi.org/10.1016/j.xpro.2023.102060 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Gurdo, Nicolás
Taylor Parkins, Shannara Kayleigh
Fricano, Martina
Wulff, Tune
Nielsen, Lars Keld
Nikel, Pablo Iván
Protocol for absolute quantification of proteins in Gram-negative bacteria based on QconCAT-based labeled peptides
title Protocol for absolute quantification of proteins in Gram-negative bacteria based on QconCAT-based labeled peptides
title_full Protocol for absolute quantification of proteins in Gram-negative bacteria based on QconCAT-based labeled peptides
title_fullStr Protocol for absolute quantification of proteins in Gram-negative bacteria based on QconCAT-based labeled peptides
title_full_unstemmed Protocol for absolute quantification of proteins in Gram-negative bacteria based on QconCAT-based labeled peptides
title_short Protocol for absolute quantification of proteins in Gram-negative bacteria based on QconCAT-based labeled peptides
title_sort protocol for absolute quantification of proteins in gram-negative bacteria based on qconcat-based labeled peptides
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9881405/
https://www.ncbi.nlm.nih.gov/pubmed/36853682
http://dx.doi.org/10.1016/j.xpro.2023.102060
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