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The mammalian-membrane two-hybrid assay (MaMTH) for probing membrane-protein interactions in human cells.
Petschnigg, Julia; Groisman, Bella; Kotlyar, Max; Taipale, Mikko; Zheng, Yong; Kurat, Christoph F; Sayad, Azin; Sierra, J Rafael; Mattiazzi Usaj, Mojca; Snider, Jamie; Nachman, Alex; Krykbaeva, Irina; Tsao, Ming-Sound; Moffat, Jason; Pawson, Tony; Lindquist, Susan; Jurisica, Igor; Stagljar, Igor.
Afiliação
  • Petschnigg J; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
  • Groisman B; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
  • Kotlyar M; Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, Ontario, Canada.
  • Taipale M; Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.
  • Zheng Y; Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
  • Kurat CF; 1] Donnelly Centre, University of Toronto, Toronto, Ontario, Canada. [2].
  • Sayad A; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
  • Sierra JR; Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, Ontario, Canada.
  • Mattiazzi Usaj M; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
  • Snider J; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
  • Nachman A; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
  • Krykbaeva I; Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.
  • Tsao MS; 1] Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, Ontario, Canada. [2] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada. [3] Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Cana
  • Moffat J; 1] Donnelly Centre, University of Toronto, Toronto, Ontario, Canada. [2] Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
  • Pawson T; 1] Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada. [2].
  • Lindquist S; 1] Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA. [2] Howard Hughes Medical Institute, Cambridge, Massachusetts, USA. [3] Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
  • Jurisica I; 1] Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, Ontario, Canada. [2] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada. [3] Department of Computer Science, University of Toronto, Toronto, Ontario, Canada.
  • Stagljar I; 1] Donnelly Centre, University of Toronto, Toronto, Ontario, Canada. [2] Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada. [3] Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
Nat Methods ; 11(5): 585-92, 2014 May.
Article em En | MEDLINE | ID: mdl-24658140
ABSTRACT
Cell signaling, one of key processes in both normal cellular function and disease, is coordinated by numerous interactions between membrane proteins that change in response to stimuli. We present a split ubiquitin-based method for detection of integral membrane protein-protein interactions (PPIs) in human cells, termed mammalian-membrane two-hybrid assay (MaMTH). We show that this technology detects stimulus (hormone or agonist)-dependent and phosphorylation-dependent PPIs. MaMTH can detect changes in PPIs conferred by mutations such as those in oncogenic ErbB receptor variants or by treatment with drugs such as the tyrosine kinase inhibitor erlotinib. Using MaMTH as a screening assay, we identified CRKII as an interactor of oncogenic EGFR(L858R) and showed that CRKII promotes persistent activation of aberrant signaling in non-small cell lung cancer cells. MaMTH is a powerful tool for investigating the dynamic interactomes of human integral membrane proteins.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Membrana Celular / Técnicas do Sistema de Duplo-Híbrido / Mapeamento de Interação de Proteínas Tipo de estudo: Prognostic_studies Limite: Humans Idioma: En Ano de publicação: 2014 Tipo de documento: Article
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Membrana Celular / Técnicas do Sistema de Duplo-Híbrido / Mapeamento de Interação de Proteínas Tipo de estudo: Prognostic_studies Limite: Humans Idioma: En Ano de publicação: 2014 Tipo de documento: Article