Dataset Information

Quantitative phosphoproteomic for CDKL5 substrates

ABSTRACT: Global phosphoproteomic screen to identify the first cellular substrates of CDKL5. CDKL5 knock-out U2OS cells and CDKL5 wt U2OS cells were generated for the TMT-based phosphoproteomic. Thi leads to the identification and further validation of several phosphopetides of MAP1S, CEP131 and CDKL5 itself. The phosphoproteomic analysis allowed the identification of the first cellular substrates for CDKL5 kinase.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

DISEASE(S): Brain Disease

SUBMITTER: Matthias Trost

LAB HEAD: Matthias Trost

PROVIDER: PXD009374 | Pride | 2018-09-24

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20160915_ppTMT_BRp_cdk5L_FR1.raw	Raw
	20160915_ppTMT_BRp_cdk5L_FR10.raw	Raw
	20160915_ppTMT_BRp_cdk5L_FR11.raw	Raw
	20160915_ppTMT_BRp_cdk5L_FR12.raw	Raw
	20160915_ppTMT_BRp_cdk5L_FR2.raw	Raw

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Publications

Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase.

Muñoz Ivan M IM Morgan Michael E ME Peltier Julien J Weiland Florian F Gregorczyk Mateusz M Brown Fiona Cm FC Macartney Thomas T Toth Rachel R Trost Matthias M Rouse John J

The EMBO journal 20180928 24

Mutations in the gene encoding the protein kinase CDKL5 cause a debilitating neurodevelopmental disease termed CDKL5 disorder. The impact of these mutations on CDKL5 function is poorly understood because the substrates and cellular processes controlled by CDKL5 are unclear. Here, we describe a quantitative phosphoproteomic screening which identified MAP1S, CEP131 and DLG5-regulators of microtubule and centrosome function-as cellular substrates of CDKL5. Antibodies against MAP1S phospho-Ser<sup>9 ...[more]

PMID: 30266825

Similar Datasets

Project description:Background: Mutations in the X-linked gene cyclin-dependent kinase-like 5 (CDKL5) cause a severe neurological disorder characterised by early-onset epileptic seizures, autism and intellectual disability (ID). Impaired hippocampal function has been implicated in other models of monogenic forms of autism spectrum disorders and ID and is often linked to epilepsy and behavioural abnormalities. Many individuals with CDKL5 deficiency disorder (CDD) have null mutations and complete loss of CDKL5 protein, therefore in the current study we used a Cdkl5-/y rat model to elucidate the impact of CDKL5 loss on cellular excitability and synaptic function of CA1 pyramidal cells (PCs). We hypothesised abnormal pre and/or post synaptic function underlie the enhanced LTP we observe in the hippocampus of Cdkl5-/y rats. Methods: To allow cross-species comparisons of phenotypes associated with the loss of CDKL5, we generated a loss of function mutation in exon 5 of the rat Cdkl5 gene and assessed the impact of the loss of CDLK5 using a combination of extracellular and whole-cell electrophysiological recordings, biochemistry, and histology. Results: Our results indicate that CA1 hippocampal LTP is enhanced in slices prepared from juvenile, but not adult, Cdkl5-/y rats. Enhanced LTP does not result from changes in NMDA receptor function or subunit expression as these remain unaltered throughout development. Furthermore, Ca2+ permeable AMPA receptor mediated currents are unchanged in Cdkl5-/y rats. We observe reduced mEPSC frequency accompanied by increased spine density in basal dendrites of CA1 PCs, however we find no evidence supporting an increase in silent synapses when assessed using a minimal stimulation protocol in slices. Additionally, we found no change in paired-pulse ratio, consistent with normal release probability at Schaffer collateral to CA1 PC synapses. Conclusions: Our data indicate a role for CDKL5 in hippocampal synaptic function and raise the possibility that altered intracellular signalling rather than synaptic deficits contribute to the altered plasticity. Limitations: This study has focussed on the electrophysiological and anatomical properties of hippocampal CA1 PCs across early postnatal development. Studies involving other brains regions, older animals and behavioural phenotypes associated with the loss of CDKL5 are needed to understand the pathophysiology of CDD.