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Proteomics-based id...
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Grimsby, SusanneUppsala universitet,Ludwiginstitutet för cancerforskning
(author)
Proteomics-based identification of proteins interacting with Smad3 : SREBP-2 forms a complex with Smad3 and inhibits its transcriptional activity
- Article/chapterEnglish2004
Publisher, publication year, extent ...
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2004-10-07
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Wiley,2004
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printrdacarrier
Numbers
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LIBRIS-ID:oai:DiVA.org:uu-72965
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https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-72965URI
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https://doi.org/10.1016/j.febslet.2004.09.069DOI
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http://kipublications.ki.se/Default.aspx?queryparsed=id:110654603URI
Supplementary language notes
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Language:English
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Summary in:English &language:-1_t
Part of subdatabase
Classification
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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Smad3 is an important component of transforming growth factor-beta (TGFbeta) intracellular signalling. To identify novel interacting proteins of Smad3, we performed pull-down assays with Smad3 constructs fused to glutathione-S-transferase. Proteins which formed complexes with these constructs were analyzed by two-dimensional gel electrophoresis, and were identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. We identified 14 proteins interacting with the Smad3 construct lacking the N-terminal Mad homology domain 1 (MH1), and 12 proteins interacting with the construct lacking the C-terminal MH2 domain. Proteins involved in signalling processes, in metabolism regulation, novel proteins, and components of cytoskeleton form four groups of interacting proteins. Interactions of AGP7, sex-determining region Y protein, actin beta and sterol regulatory element binding protein-2 (SREBP-2) proteins with Smad3 constructs were confirmed by immunoblotting with specific antibodies. Interaction of Smad3 with SREBP-2 was also confirmed by co-immunoprecipitation of myc-Smad3 and Flag-SREBP-2 upon expression in mammalian cells. We found that SREBP-2 inhibited the transcriptional activity of Smad3 in luciferase reporter assays.
Subject headings and genre
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Animals
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Cell Line
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DNA-Binding Proteins/*metabolism
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Electrophoresis; Gel; Two-Dimensional
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Humans
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Protein Binding
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Proteome
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Research Support; Non-U.S. Gov't
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Spectrometry; Mass; Matrix-Assisted Laser Desorption-Ionization
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Trans-Activators/*metabolism
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Transcription Factors/*metabolism
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Transcription; Genetic
Added entries (persons, corporate bodies, meetings, titles ...)
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Jaensson, HannaUppsala universitet,Ludwiginstitutet för cancerforskning
(author)
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Dubrovska, AnnaUppsala universitet,Ludwiginstitutet för cancerforskning(Swepub:uu)andub498
(author)
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Lomnytska, MartaUppsala universitet,Ludwiginstitutet för cancerforskning(Swepub:uu)marlo829
(author)
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Hellman, UlfUppsala universitet,Ludwiginstitutet för cancerforskning(Swepub:uu)ulfhm
(author)
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Souchelnytskyi, SerhiyKarolinska Institutet,Uppsala universitet,Ludwiginstitutet för cancerforskning(Swepub:uu)sergsouc
(author)
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Uppsala universitetLudwiginstitutet för cancerforskning
(creator_code:org_t)
Related titles
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In:FEBS Letters: Wiley577:1-2, s. 93-1000014-57931873-3468
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