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Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Viertler, M; Schittmayer, M; Birner-Gruenberger, R.
Activity based subcellular resolution imaging of lipases.
Bioorg Med Chem. 2012; 20(2):628-632
Doi: 10.1016/j.bmc.2011.04.018
Web of Science
PubMed
FullText
FullText_MUG
- Führende Autor*innen der Med Uni Graz
- Birner-Grünberger Ruth
- Schittmayer-Schantl Matthias
- Viertler Martin
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- Abstract:
- Lipases play a key role in whole body energy homeostasis. Dysregulation of lipolytic activities affects lipid absorption, mobilization, and transport, and is causative for lipid-related diseases. Regulation of enzymes involved in lipid metabolism is governed by a complex network of protein-protein and protein-small molecule interactions. Thus these enzymes have to be studied under the physiologically most relevant conditions, that is, in vivo. Our latest generation of activity based probes designed for capturing of lipases employs bioorthogonal chemical linker groups, which are membrane permeable and thus allow studying protein activity in living cells. Another advantage is the virtually unlimited choice of reporter tags. Here we report on a novel method combining in vivo activity based labeling of lipases with in situ detection of lipolytic activities by on slide click chemistry and imaging by fluorescence microscopy. We demonstrate that cytosolic as well as organelle resident lipases are specifically labeled in intact living cells. This method will shed light on the (sub)cellular localization of lipolytic proteomes of cells and tissues in health and disease directly at enzymatic activity level without the need of prior knowledge of the identities of the responsible enzymes or dependence on the availability of specific antibodies.
- Find related publications in this database (using NLM MeSH Indexing)
- Alkynes - chemistry
- Animals -
- Azides - chemistry
- COS Cells -
- Catalysis -
- Cercopithecus aethiops -
- Click Chemistry -
- Copper - chemistry
- Lipase - chemistry Lipase - metabolism
- Lipid Metabolism -
- Microscopy, Fluorescence -
- Organophosphonates - chemical synthesis Organophosphonates - chemistry
- Proteome - analysis
- Find related publications in this database (Keywords)
- Activity based proteomics
- On-slide click chemistry
- Activity based cell imaging
- Subcellular lipase localization