التفاصيل البيبلوغرافية
| العنوان: |
Subcellular imaging of lipids and sugars using genetically encoded proximity sensors. |
| المؤلفون: |
Moore WM, Brea RJ, Knittel C, Wrightsman E, Hui B, Lou J, Ancajas CF, Best MD, Devaraj NK, Budin I |
| المصدر: |
BioRxiv : the preprint server for biology [bioRxiv] 2024 May 03. Date of Electronic Publication: 2024 May 03. |
| نوع المنشور: |
Preprint |
| اللغة: |
English |
| بيانات الدورية: |
Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE |
| مستخلص: |
Live cell imaging of lipids and other metabolites is a long-standing challenge in cell biology. Bioorthogonal labeling tools allow for the conjugation of fluorophores to several phospholipid classes, but cannot discern their trafficking between adjacent organelles or asymmetry across individual membrane leaflets. Here we present fluorogen-activating coincidence sensing (FACES), a chemogenetic tool capable of quantitatively imaging subcellular lipid pools and reporting their transbilayer orientation in living cells. FACES combines bioorthogonal chemistry with genetically encoded fluorogen-activating proteins (FAPs) for reversible proximity sensing of conjugated molecules. We first validate this approach for quantifying discrete phosphatidylcholine pools in the ER and mitochondria that are trafficked by lipid transfer proteins. We then show that transmembrane domain-containing FAPs can be used to reveal the membrane asymmetry of multiple lipid classes that are generated in the trans-Golgi network. Lastly, we demonstrate that FACES is a generalizable tool for subcellular bioorthogonal imaging by measuring changes in mitochondrial N -acetylhexosamine levels. These results demonstrate the use of fluorogenic tags for spatially-defined molecular imaging. |
| معلومات مُعتمدة: |
R35 GM142960 United States GM NIGMS NIH HHS |
| تواريخ الأحداث: |
Date Created: 20240515 Latest Revision: 20240615 |
| رمز التحديث: |
20240615 |
| مُعرف محوري في PubMed: |
PMC11092643 |
| DOI: |
10.1101/2024.05.01.592120 |
| PMID: |
38746395 |
| قاعدة البيانات: |
MEDLINE |
