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PeerJ. 2018 Apr 9;6:e4599. doi: 10.7717/peerj.4599. eCollection 2018.

Phosphoproteomic insights into processes influenced by the kinase-like protein DIA1/C3orf58.

Author information

1
Department of Experimental Design and Bioinformatics, Faculty of Agriculture and Biology, Warsaw University of Life Sciences, Warszawa, Poland.
2
International Institute of Molecular and Cellular Biology, Warszawa, Poland.
3
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland.
4
Current affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
5
Department of Translational Medicine, Clinical Sciences, Lund University, Lund, Sweden.

Abstract

Many kinases are still 'orphans,' which means knowledge about their substrates, and often also about the processes they regulate, is lacking. Here, DIA1/C3orf58, a member of a novel predicted kinase-like family, is shown to be present in the endoplasmic reticulum and to influence trafficking via the secretory pathway. Subsequently, DIA1 is subjected to phosphoproteomics analysis to cast light on its signalling pathways. A liquid chromatography-tandem mass spectrometry proteomic approach with phosphopeptide enrichment is applied to membrane fractions of DIA1-overexpressing and control HEK293T cells, and phosphosites dependent on the presence of DIA1 are elucidated. Most of these phosphosites belonged to CK2- and proline-directed kinase types. In parallel, the proteomics of proteins immunoprecipitated with DIA1 reported its probable interactors. This pilot study provides the basis for deeper studies of DIA1 signalling.

KEYWORDS:

Mass spectrometry; Novel kinases; Phosphoproteomics; Secretory pathway; Signalling

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