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The temporal profile of activity-dependent presynaptic phospho-signalling reveals a PP1-dependent reduction in vesicle release

ABSTRACT: Depolarization of presynaptic terminals stimulates calcium influx, which evokes neurotransmitter release and activates phosphorylation-based signalling. Here, we present the first global temporal profile of presynaptic proteins undergoing post-stimulus phospho-signalling to reveal long-lasting changes, key substrates and master regulators. A total of 5,715 unique phosphopeptides from 1,817 proteins were profiled and 1,917 phosphopeptides had activity-dependent phosphorylation sites. A new computational method, KinSwing, combining protein kinase substrate prediction and activity across time, enabled the prediction of effector protein kinase activity. CaMKII responded rapidly to depolarization. MAPK, CDK5 and GSK3β had a post-stimulus role in compensating for initial dephosphorylation. Despite this, the post-stimulus period was dominated by down-regulation of phosphorylation and was exacerbated by conditions stimulating increased calcium influx. Down-regulated phosphorylation correlated with perturbed phospho-signalling to protein phosphatase 1 (PP1) regulatory molecules and reduced glutamate and fluorescent dye release. Inhibition PP1 prevented reduction in dye release, identifying PP1 as a major signalling target and mediator of post-stimulus presynaptic phospho-signalling.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Brain

SUBMITTER: Kasper Engholm-Keller

LAB HEAD: Mark Graham

PROVIDER: PXD008495 | Pride | 2019-01-24

REPOSITORIES: Pride

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			Action	DRS
	evidence.txt	Txt
	mono_HILIC_frac10_rep1_20_1.raw	Raw
	mono_HILIC_frac10_rep1_20_2.raw	Raw
	mono_HILIC_frac10_rep1_80_1.raw	Raw
	mono_HILIC_frac10_rep1_80_2.raw	Raw

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The temporal profile of activity-dependent presynaptic phospho-signalling reveals long lasting patterns of post-stimulus regulation

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