MassIVE MSV000086462

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Arabidopsis thaliana whole-proteome BIN2 kinase assay

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Description

Finely ground leaf tissue of wild-type columbia-0 ecotype was used. Protein extraction was performed in buffer-saturated phenol and high sucrose buffer. Extracted proteins were incubated with purified recombinant GST or GST-BIN2 kinase. Filter-assisted sample preparation (FASP) columns in presence of 8M urea were used to further purify proteins. Purified proteins were reduced with TCEP, alkylated with iodoacetamide and digested into peptides with trypsin and lys-C. Digested peptides were desalted with c18 columns and labelled with 11-plex Tandem Mass Tag (TMT11) reagents. Phosphoenrichment was performed in a two-steps tandem pipeline: High-Select TiO2 Phosphopeptide Enrichment kit (Thermo Scientific) was used first and High Select Fe-NTA Phosphoptide Enrichment kit (Thermo Scientific) was used on the flow-through obtained from first enrichment. [doi:10.25345/C5ZN3W] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: BIN2 brassinosteroids kinase

Contact

Principal Investigators:
(in alphabetical order) 		Justin Walley, Iowa State University, US
Submitting User: chrisfm

Publications

Montes C, Wang P, Liao CY, Nolan TM, Song G, Clark NM, Elmore JM, Guo H, Bassham DC, Yin Y, Walley JW.
Integration of multi-omics data reveals interplay between brassinosteroid and Target of Rapamycin Complex signaling in Arabidopsis.
New Phytol. 2022 Nov;236(3):893-910. Epub 2022 Aug 11.

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