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Utilization of Two-Photon FRET to Monitor SREBP Homodimer and Heterodimer Formation In Living Cells

机译：利用双光子FRET监测活细胞中SREBP均聚物和异二聚体的形成

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摘要

Key players in cholesterol regulation are the members of a family of transcription factors known as the Sterol Regulatory Binding Proteins or SREBPs. The cellular redundancy of these proteins is under investigation, and our findings suggest that where these proteins reside may provide evidence for differences in the molecular dynamics of their transcriptional activity. Specifically, we have found that GFP-tagged SREBP-2 in contrast to SREBP-1 resides in discrete nuclear foci. To further explore functional differences between SREBP-1 and SREBP-2 we have developed an approach to monitor hetero- and homodimer formation by two-photon imaging and spectroscopy of fluorescence resonance energy transfer (TPIS-FRET). TPIS-FRET results will be presented. Collectively, these findings support the possibility that differences in function between SREBP family members may be governed by their localization within the cell.
机译：胆固醇调节的关键参与者是转录因子家族的成员，这些转录因子称为Sterol调节结合蛋白或SREBPs。这些蛋白质的细胞冗余性正在研究中，我们的发现表明这些蛋白质所处的位置可能为其转录活性的分子动力学差异提供证据。具体而言，我们发现与SREBP-1相反，带有GFP标签的SREBP-2位于离散的核灶中。为了进一步探索SREBP-1和SREBP-2之间的功能差异，我们开发了一种通过双光子成像和荧光共振能量转移光谱（TPIS-FRET）监测异二聚体和同二聚体形成的方法。将显示TPIS-FRET结果。总的来说，这些发现支持SREBP家族成员之间功能差异可能受其在细胞内的定位支配的可能性。

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