PUBLICATION
A Photo-clickable ATP-Mimetic Reveals Nucleotide Interactors in the Membrane Proteome
- Authors
- Jelcic, M., Wang, K., Hui, K.L., Cai, X.C., Enyedi, B., Luo, M., Niethammer, P.
- ID
- ZDB-PUB-210707-16
- Date
- 2020
- Source
- Cell chemical biology 27: 1073-1083.e12 (Journal)
- Registered Authors
- Enyedi, Balázs, Niethammer, Philipp
- Keywords
- ATP, ATP photoaffinity probe, calcium signaling, chemical proteomics, photoaffinity labeling, purinergic signaling, quantitative proteomics, target discovery
- MeSH Terms
-
- Calcium Signaling
- Larva/metabolism
- Ultraviolet Rays
- Calmodulin/genetics
- Calmodulin/metabolism
- Cell Membrane/chemistry
- Cell Membrane/metabolism*
- Humans
- Tandem Mass Spectrometry
- Adenosine Triphosphate/analogs & derivatives
- Adenosine Triphosphate/chemical synthesis
- Adenosine Triphosphate/metabolism*
- Isotope Labeling
- Proteome/analysis*
- Proteome/metabolism
- Chromatography, High Pressure Liquid
- Animals, Genetically Modified/metabolism
- Click Chemistry
- Animals
- Fluorescent Dyes/chemistry
- Amino Acids/chemistry
- Amino Acids/metabolism
- Zebrafish/growth & development
- Zebrafish/metabolism
- Cell Line, Tumor
- Optical Imaging
- PubMed
- 32521230 Full text @ Cell Chem Biol
Citation
Jelcic, M., Wang, K., Hui, K.L., Cai, X.C., Enyedi, B., Luo, M., Niethammer, P. (2020) A Photo-clickable ATP-Mimetic Reveals Nucleotide Interactors in the Membrane Proteome. Cell chemical biology. 27:1073-1083.e12.
Abstract
ATP is an important energy metabolite and allosteric signal in health and disease. ATP-interacting proteins, such as P2 receptors, control inflammation, cell death, migration, and wound healing. However, identification of allosteric ATP sites remains challenging, and our current inventory of ATP-controlled pathways is likely incomplete. Here, we develop and verify mipATP as a minimally invasive photoaffinity probe for ATP-interacting proteins. Its N6 functionalization allows target enrichment by UV crosslinking and conjugation to reporter tags by "click" chemistry. The additions are compact, allowing mipATP to completely retain the calcium signaling responses of native ATP in vitro and in vivo. mipATP specifically enriched for known nucleotide binders in A549 cell lysates and membrane fractions. In addition, it retrieved unannotated ATP interactors, such as the FAS receptor, CD44, and various SLC transporters. Thus, mipATP is a promising tool to identify allosteric ATP sites in the proteome.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping