Identification of novel Smad2 and Smad3 associated proteins in response to TGF-beta1.

Journal of Cellular Biochemistry
Kimberly A BrownHarold L Moses

Abstract

Transforming growth factor-beta 1 (TGF-beta1) is an important growth inhibitor of epithelial cells and insensitivity to this cytokine results in uncontrolled cell proliferation and can contribute to tumorigenesis. TGF-beta1 signals through the TGF-beta type I and type II receptors, and activates the Smad pathway via phosphorylation of Smad2 and Smad3. Since little is known about the selective activation of Smad2 versus Smad3, we set out to identify novel Smad2 and Smad3 interacting proteins in epithelial cells. A non-transformed human cell line was transduced with Myc-His(6)-Smad2 or Myc-His(6)-Smad3-expressing retrovirus and was treated with TGF-beta1. Myc-His(6)-Smad2 or Myc-His(6)-Smad3 was purified by tandem affinity purification, eluates were subject to SDS-PAGE and Colloidal Blue staining, and select protein bands were digested with trypsin. The resulting tryptic peptides were analyzed by liquid chromatography (LC) and tandem mass spectrometry (MS/MS) and the SEQUEST algorithm was employed to identify proteins in the bands. A number of proteins that are known to interact with Smad2 or Smad3 were detected in the eluates. In addition, a number of putative novel Smad2 and Smad3 associated proteins were identified that have f...Continue Reading

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Citations

Feb 17, 2010·Biotechnology and Applied Biochemistry·Yifeng Li
Apr 5, 2017·The Journal of Clinical Investigation·Zhe YingMengfeng Li
Dec 1, 2017·Cell Communication and Signaling : CCS·Eva María García-VizcaínoFrancisco José Nicolás
Oct 14, 2017·Rheumatology·Mesias PedrozaSandeep K Agarwal
Jan 25, 2020·Molecular Neurobiology·Maria A KashermanMichael Piper
Apr 24, 2009·Expert Review of Proteomics·Maria MontiPiero Pucci
Nov 24, 2016·PloS One·Estibaliz AlvaradoRobert M Nissen
Oct 4, 2017·Scientific Reports·Rabab SharifDimitrios Karamichos
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Jul 1, 2020·Cancer Gene Therapy·Jane AntonyChiara Recchi
Feb 7, 2021·Angiogenesis·Clare E Benson, Laura Southgate
Oct 3, 2009·Cellular Signalling·Yuki Miyamoto, Junji Yamauchi

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