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Calcium-dependent proximal biotinylation via Cal-ID

ABSTRACT: Calcium ions serve as key intracellular signals. Local, transient increases in calcium concentrations can activate calcium sensor proteins that in turn trigger downstream effectors. In neurons, such calcium transients trigger pre-synaptic vesicle release and mediate post-synaptic plasticity. It is challenging to capture the molecular events associated with these localized and ephemeral calcium signals, however. Here we report the development of an engineered biotin ligase that combines the power of genetically encoded calcium indicators with protein proximity labeling. The enzyme, Cal-ID, biotinylates nearby proteins in response to elevated local calcium levels. The biotinylated proteins can be visualized by microscopy and identified via mass spectrometry. Cal-ID mass spectrometry applied to HEK293T cells identified cell cycle-dependent calcium signaling microdomains at centrosomes. Our results from mouse primary neurons indicated active calcium signaling near calcium extrusion sites on the plasma membrane. Therefore, we propose Cal-ID as a biochemical recorder of calcium signaling in living cells.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

SUBMITTER: J Wren Kim

LAB HEAD: Nicholas Ingolia

PROVIDER: PXD033244 | Pride | 2024-02-04

REPOSITORIES: Pride

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			Action	DRS
	JWK_Cal-ID_HEK_TMT.raw	Raw
	JWK_Cal-ID_HEK_results.xlsx	Xlsx
	JWK_Cal-ID_mCN_TMT.raw	Raw
	JWK_Cal-ID_mCN_results.xlsx	Xlsx
	checksum.txt	Txt

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