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| Johannes A. Schmid; Andreas Birbach. |
| IKK2 is one of the most crucial signaling kinases for activation of the transcription factor NF-kappa B. Since many NF-kappa B activating pathways converge at the level of IKK2, we searched for interaction partners of this kinase using the C-terminal part (aa 466-756) as bait in a yeast two-hybrid system. We identified the N-terminal part (aa 1-228) of the TNF-receptor associated factor TRAF1 as putative interaction partner, which was subsequently confirmed in mammalian cells by coimmunoprecipitation experiments. However, this interaction seemed weaker than the interaction between TRAF1 and TRAF2, an important activating adapter molecule of NF-kappa B signaling indicating that relative levels of IKK2, TRAF1 and TRAF2 might be important for the final... |
| Tipo: Manuscript |
Palavras-chave: Immunology; Molecular Cell Biology. |
| Ano: 2010 |
URL: http://precedings.nature.com/documents/4153/version/1 |
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| Johannes A. Schmid. |
| The use of spectrally distinct variants of green fluorescent protein (GFP) such as cyan or yellow mutants (CFP and YFP, respectively) is very common in all different fields of life sciences, e.g. for marking and tracing of specific proteins or cells or to determine protein interactions. In the later case, the quantum physical phenomenon of fluorescence resonance energy transfer (FRET) is visualized by specific microscopy techniques. When we applied a commonly used FRET microscopy technique - the increase in CFP-fluorescence after bleaching of YFP, we noticed that it worked well for live cells, but that most of the FRET-signal was lost in fixed cells mounted in commercial microscopy mounting fluids. Subsequently, we could show that CFP bleached much faster... |
| Tipo: Manuscript |
Palavras-chave: Biotechnology; Molecular Cell Biology. |
| Ano: 2010 |
URL: http://precedings.nature.com/documents/4517/version/1 |
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| Kalsoom A. Sughra; Andreas Birbach; Rainer de Martin; Johannes A. Schmid. |
| *Background*
I-kappa B kinase 2 (IKK2 or IKK-beta) is one of the most crucial signaling kinases for activation of NF-kappa B, a transcription factor that is important for inflammation, cell survival and differentiation. Since many NF-kappa B activating pathways converge at the level of IKK2, molecular interactions of this kinase are pivotal for regulation of NF-kappa B signaling.

*Methodology/Principal Findings*
We searched for proteins interacting with IKK2 using the C-terminal part (amino acids 466-756) as bait in a yeast two-hybrid system and identified the N-terminal part (amino acids 1-228) of the TNF-receptor associated factor TRAF1 as putative interaction partner. The interaction... |
| Tipo: Manuscript |
Palavras-chave: Immunology; Molecular Cell Biology. |
| Ano: 2010 |
URL: http://precedings.nature.com/documents/4153/version/2 |
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