Dataset Information

Identification of N-terminally arginylated proteins from the moss Physcomitrella patens

ABSTRACT: Protein arginylation is a post-translational modification at the N-terminus of proteins and is crucial for viability and physiology in higher eukaryotes. The lack of arginylation causes severe developmental defects in plants and embryo lethality in Drosophila and in mice. Although several studies investigated impact and function of the responsible enzyme, the arginyl-tRNA protein transferase (ATE) in plants, identification of arginylated proteins by mass spectrometry was not hitherto achieved. In the present study, we report the identification of targets and interaction partners of ATE in the model plant Physcomitrella patens by mass spectrometry employing two different immuno-affinity strategies and a recently established transgenic ATE:GUS reporter line (Schuessele et al., 2015 New Phytol., DOI: 10.1111/nph.13656). Here we use a commercially available antibody against the fused reporter protein (GUS) to pull down ATE and its interacting proteins and validate the in vivo interaction with a class I small heatshock protein via Förster resonance energy transfer (FRET). Additionally, we apply and modify a method that already successfully identified arginylated proteins from mouse proteomes by using custom-made antibodies specific for N-terminal arginine. As a result, we identify four arginylated proteins from Physcomitrella patens with high confidence.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Physcomitrella Patens Subsp. Patens (moss)

TISSUE(S): Gametophore

SUBMITTER: Ralf Reski

LAB HEAD: Ralf Reski

PROVIDER: PXD003232 | Pride | 2016-04-21

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	2013-10-02_velos_5130.raw	Raw
	2013-10-02_velos_5131.raw	Raw
	2013-10-02_velos_5132.raw	Raw
	2013-10-02_velos_5133.raw	Raw
	2013-10-02_velos_5134.raw	Raw

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Publications

Identification of Targets and Interaction Partners of Arginyl-tRNA Protein Transferase in the Moss Physcomitrella patens.

Hoernstein Sebastian N W SN Mueller Stefanie J SJ Fiedler Kathrin K Schuelke Marc M Vanselow Jens T JT Schuessele Christian C Lang Daniel D Nitschke Roland R Igloi Gabor L GL Schlosser Andreas A Reski Ralf R

Molecular & cellular proteomics : MCP 20160411 6

Protein arginylation is a posttranslational modification of both N-terminal amino acids of proteins and sidechain carboxylates and can be crucial for viability and physiology in higher eukaryotes. The lack of arginylation causes severe developmental defects in moss, affects the low oxygen response in Arabidopsis thaliana and is embryo lethal in Drosophila and in mice. Although several studies investigated impact and function of the responsible enzyme, the arginyl-tRNA protein transferase (ATE) i ...[more]

PMID: 27067052

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