Combining chemical and genetic approaches for development of responsive FRET-based sensor systems for protein kinases
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- 作者：Ganesh babu Manoharan ; Erki Enkvist ; Asko Uri ; asko.uri@ut.ee" class="auth_mail" title="E-mail the corresponding author
- 关键词：Protein kinase ; PIM2 ; FRET ; Phosphorescence ; Fluorescence ; Responsive probes
- 刊名：Biophysical Chemistry
- 出版年：2016
- 出版时间：April 2016
- 年：2016
- 卷：211
- 期：Complete
- 页码：39-48
- 全文大小：1714 K
文摘
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Steady-state and time-gated FRET sensor systems were developed based on singlet-singlet and triplet–singlet energy transfers.
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Triplet–singlet energy transfer between selenium-comprising heteroaromatic fragment of ARC-Lum(−) probe and TagRFP-fused PIM2
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Transport peptide nona(d-arginine) amide binds to PIM2 kinase with high affinity.
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Time-gated FRET assay using Lumi4Tb-labeled ARC probes and TagRFP-fused PIM2
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PIM2 concentration in cell lysate was determined using ARC-Lum probes.