SLAPSHOT reveals rapid dynamics of extracellularly exposed proteome in response to calcium-activated plasma membrane phospholipid scrambling.

Autor: Tuomivaara ST; Department of Laboratory Medicine, University of California, San Francisco, CA, USA.; Department of Obstetrics, Gynecology & Reproductive Sciences and Sandler-Moore Mass Spectrometry Core Facility, University of California, San Francisco, CA, USA., Teo CF; Howard Hughes Medical Institute, University of California, San Francisco, CA, USA.; Department of Physiology, University of California, San Francisco, CA, USA., Jan YN; Howard Hughes Medical Institute, University of California, San Francisco, CA, USA.; Department of Physiology, University of California, San Francisco, CA, USA.; Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA., Wiita AP; Department of Laboratory Medicine, University of California, San Francisco, CA, USA. arun.wiita@ucsf.edu.; Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA, USA. arun.wiita@ucsf.edu.; Chan Zuckerberg Biohub San Francisco, San Francisco, CA, USA. arun.wiita@ucsf.edu., Jan LY; Howard Hughes Medical Institute, University of California, San Francisco, CA, USA. lily.jan@ucsf.edu.; Department of Physiology, University of California, San Francisco, CA, USA. lily.jan@ucsf.edu.; Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA. lily.jan@ucsf.edu.
Jazyk: angličtina
Zdroj: Communications biology [Commun Biol] 2024 Aug 29; Vol. 7 (1), pp. 1060. Date of Electronic Publication: 2024 Aug 29.
DOI: 10.1038/s42003-024-06729-x
Abstrakt: To facilitate our understanding of proteome dynamics during signaling events, robust workflows affording fast time resolution without confounding factors are essential. We present Surface-exposed protein Labeling using PeroxidaSe, H 2 O 2 , and Tyramide-derivative (SLAPSHOT) to label extracellularly exposed proteins in a rapid, specific, and sensitive manner. Simple and flexible SLAPSHOT utilizes recombinant soluble APEX2 protein applied to cells, thus circumventing the engineering of tools and cells, biological perturbations, and labeling biases. We applied SLAPSHOT and quantitative proteomics to examine the TMEM16F-dependent plasma membrane remodeling in WT and TMEM16F KO cells. Time-course data ranging from 1 to 30 min of calcium stimulation revealed co-regulation of known protein families, including the integrin and ICAM families, and identified proteins known to reside in intracellular organelles as occupants of the freshly deposited extracellularly exposed membrane. Our data provide the first accounts of the immediate consequences of calcium signaling on the extracellularly exposed proteome.
(© 2024. The Author(s).)
Databáze: MEDLINE
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