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Combining RNA interference mutants and comparative proteomics to identify protein component interactions and dependencies in a eukaryotic flagellum.

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Combining RNA interference mutants and comparative proteomics to identify protein component interactions and dependencies in a eukaryotic flagellum. / Portman, Neil; Lacomble, Sylvain; Thomas, Benjamin et al.
In: Journal of Biological Chemistry, Vol. 284, No. 9, 02.2009, p. 5610-5619.

Research output: Contribution to Journal/MagazineJournal articlepeer-review

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Portman N, Lacomble S, Thomas B, McKean PG, Gull K. Combining RNA interference mutants and comparative proteomics to identify protein component interactions and dependencies in a eukaryotic flagellum. Journal of Biological Chemistry. 2009 Feb;284(9):5610-5619. doi: 10.1074/jbc.M808859200

Author

Portman, Neil ; Lacomble, Sylvain ; Thomas, Benjamin et al. / Combining RNA interference mutants and comparative proteomics to identify protein component interactions and dependencies in a eukaryotic flagellum. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 9. pp. 5610-5619.

Bibtex

@article{4fe8f00994304f958aa687dd5d843698,
title = "Combining RNA interference mutants and comparative proteomics to identify protein component interactions and dependencies in a eukaryotic flagellum.",
abstract = "Eukaryotic flagella from organisms such as Trypanosoma brucei can be isolated and their protein components identified by mass spectrometry. Here we used a comparative approach utilizing two-dimensional difference gel electrophoresis and isobaric tags for relative and absolute quantitation to reveal protein components of flagellar structures via ablation by inducible RNA interference mutation. By this approach we identified 20 novel components of the paraflagellar rod (PFR). Using epitope tagging we validated a subset of these as being present within the PFR by immunofluorescence. Bioinformatic analysis of the PFR cohort reveals a likely calcium/calmodulin regulatory/signaling linkage between some components. We extended the RNA interference mutant/comparative proteomic analysis to individual novel components of our PFR proteome, showing that the approach has the power to reveal dependences between subgroups within the cohort.",
author = "Neil Portman and Sylvain Lacomble and Benjamin Thomas and McKean, {Paul G.} and Keith Gull",
year = "2009",
month = feb,
doi = "10.1074/jbc.M808859200",
language = "English",
volume = "284",
pages = "5610--5619",
journal = "Journal of Biological Chemistry",
issn = "1083-351X",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "9",

}

RIS

TY - JOUR

T1 - Combining RNA interference mutants and comparative proteomics to identify protein component interactions and dependencies in a eukaryotic flagellum.

AU - Portman, Neil

AU - Lacomble, Sylvain

AU - Thomas, Benjamin

AU - McKean, Paul G.

AU - Gull, Keith

PY - 2009/2

Y1 - 2009/2

N2 - Eukaryotic flagella from organisms such as Trypanosoma brucei can be isolated and their protein components identified by mass spectrometry. Here we used a comparative approach utilizing two-dimensional difference gel electrophoresis and isobaric tags for relative and absolute quantitation to reveal protein components of flagellar structures via ablation by inducible RNA interference mutation. By this approach we identified 20 novel components of the paraflagellar rod (PFR). Using epitope tagging we validated a subset of these as being present within the PFR by immunofluorescence. Bioinformatic analysis of the PFR cohort reveals a likely calcium/calmodulin regulatory/signaling linkage between some components. We extended the RNA interference mutant/comparative proteomic analysis to individual novel components of our PFR proteome, showing that the approach has the power to reveal dependences between subgroups within the cohort.

AB - Eukaryotic flagella from organisms such as Trypanosoma brucei can be isolated and their protein components identified by mass spectrometry. Here we used a comparative approach utilizing two-dimensional difference gel electrophoresis and isobaric tags for relative and absolute quantitation to reveal protein components of flagellar structures via ablation by inducible RNA interference mutation. By this approach we identified 20 novel components of the paraflagellar rod (PFR). Using epitope tagging we validated a subset of these as being present within the PFR by immunofluorescence. Bioinformatic analysis of the PFR cohort reveals a likely calcium/calmodulin regulatory/signaling linkage between some components. We extended the RNA interference mutant/comparative proteomic analysis to individual novel components of our PFR proteome, showing that the approach has the power to reveal dependences between subgroups within the cohort.

UR - http://www.scopus.com/inward/record.url?scp=65549110811&partnerID=8YFLogxK

U2 - 10.1074/jbc.M808859200

DO - 10.1074/jbc.M808859200

M3 - Journal article

VL - 284

SP - 5610

EP - 5619

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 1083-351X

IS - 9

ER -