A Photo-clickable ATP-Mimetic Reveals Nucleotide Interactors in the Membrane Proteome
| Autor: | Mark Jelcic, Xiao-Chuan Cai, Philipp Niethammer, King Lam Hui, Minkui Luo, Balázs Enyedi, Ke Wang |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Proteome
Ultraviolet Rays Clinical Biochemistry Allosteric regulation Quantitative proteomics Biology 01 natural sciences Biochemistry Article Animals Genetically Modified Adenosine Triphosphate Calmodulin Tandem Mass Spectrometry Cell Line Tumor Drug Discovery Animals Humans Nucleotide Calcium Signaling Amino Acids Receptor Molecular Biology Chromatography High Pressure Liquid Zebrafish Fluorescent Dyes Calcium signaling Pharmacology chemistry.chemical_classification Photoaffinity labeling 010405 organic chemistry Cell Membrane Optical Imaging Purinergic signalling 0104 chemical sciences chemistry Isotope Labeling Larva Molecular Medicine Click Chemistry |
| Zdroj: | Cell Chem Biol Cell Chemical Biology |
| ISSN: | 2451-9456 |
| DOI: | 10.1016/j.chembiol.2020.05.010 |
| Popis: | Summary 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. |
| Databáze: | OpenAIRE |
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