SLAPSHOT reveals rapid dynamics of extracellularly exposed proteome in response to calcium-activated plasma membrane phospholipid scrambling
Abstrak
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, H2O2, 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. Surface-exposed protein Labeling using PeroxidaSe, H2O2, and Tyramide-derivative (SLAPSHOT) coupled with mass spectrometry reveals the dynamics in the extracellularly exposed protein landscape in short time scales.
Topik & Kata Kunci
Penulis (5)
S. Tuomivaara
Chin Fen Teo
Y. Jan
Arun P. Wiita
L. Jan
Akses Cepat
- Tahun Terbit
- 2024
- Bahasa
- en
- Total Sitasi
- 4×
- Sumber Database
- Semantic Scholar
- DOI
- 10.1038/s42003-024-06729-x
- Akses
- Open Access ✓