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Screening a Resourc...
Screening a Resource of Recombinant Protein Fragments for Targeted Proteomics
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- Edfors, Fredrik (författare)
- KTH,Systembiologi,Science for Life Laboratory, SciLifeLab
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- Forsström, Björn (författare)
- KTH,Systembiologi,Science for Life Laboratory, SciLifeLab
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- Vunk, Helian (författare)
- KTH,Systembiologi,Science for Life Laboratory, SciLifeLab
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- Kotol, David (författare)
- KTH,Systembiologi,Science for Life Laboratory, SciLifeLab
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- Fredolini, Claudia (författare)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab
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- Maddalo, Gianluca (författare)
- KTH,Systembiologi,Science for Life Laboratory, SciLifeLab
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- Svensson, Anne-Sophie (författare)
- KTH
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- Boström, Tove (författare)
- KTH
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- Tegel, Hanna (författare)
- KTH
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- Nilsson, Peter (författare)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab
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- Schwenk, Jochen M. (författare)
- KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab
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- Uhlén, Mathias (författare)
- KTH,Systembiologi,Science for Life Laboratory, SciLifeLab,Karolinska Inst, Dept Neurosci, SE-17165 Solna, Sweden.;Tech Univ Denmark, Novo Nordisk Fdn Ctr Biosustainabil, DK-2970 Horsholm, Denmark.
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(creator_code:org_t)
- 2019-05-16
- 2019
- Engelska.
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Ingår i: Journal of Proteome Research. - : American Chemical Society (ACS). - 1535-3893 .- 1535-3907. ; 18:7, s. 2706-2718
- Relaterad länk:
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https://www.biorxiv....
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https://urn.kb.se/re...
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https://doi.org/10.1...
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http://kipublication...
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Abstract
Ämnesord
Stäng
- The availability of proteomics resources hosting protein and peptide standards, as well as the data describing their analytical performances, will continue to enhance our current capabilities to develop targeted proteomics methods for quantitative biology. This study describes the analysis of a resource of 26,840 individually purified recombinant protein fragments corresponding to more than 16,000 human protein-coding genes. The resource was screened to identify proteotypic peptides suitable for targeted proteomics efforts, and we report LC-MS/MS assay coordinates for more than 25,000 proteotypic peptides, corresponding to more than 10,000 unique proteins. Additionally, peptide formation and digestion kinetics were, for a subset of the standards, monitored using a time-course protocol involving parallel digestion of isotope-labeled recombinant protein standards and endogenous human plasma proteins. We show that the strategy by adding isotope-labeled recombinant proteins before trypsin digestion enables short digestion protocols (<= 60 min) with robust quantitative precision. In a proof-of-concept study, we quantified 23 proteins in human plasma using assay parameters defined in our study and used the standards to describe distinct clusters of individuals linked to different levels of LPA, APOE, SERPINAS, and TFRC. In summary, we describe the use and utility of a resource of recombinant proteins to identify proteotypic peptides useful for targeted proteomics assay development.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- targeted proteomics
- stable isotope standards
- mass spectrometry
- protein quantification
- recombinant proteins
- protein fragment
- trypsin digestion
- spectral library
- assay generation
- peptide formation
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- ref (ämneskategori)
- art (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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Edfors, Fredrik
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Forsström, Björn
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Vunk, Helian
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Kotol, David
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Fredolini, Claud ...
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Maddalo, Gianluc ...
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visa fler...
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Svensson, Anne-S ...
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Boström, Tove
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Tegel, Hanna
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Nilsson, Peter
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Schwenk, Jochen ...
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Uhlén, Mathias
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- Om ämnet
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- NATURVETENSKAP
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NATURVETENSKAP
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och Biologi
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och Biokemi och mole ...
- Artiklar i publikationen
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Journal of Prote ...
- Av lärosätet
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Kungliga Tekniska Högskolan
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Karolinska Institutet