MassIVE MSV000094195

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Multivalent Coiled-Coil Interactions as a Conserved Design Principle for Centrosome Assembly

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Description

Purified GFP-CDK5RAP2 (in Storage Buffer; 25 mM HEPES, pH 7.4, 500 mM KCl, 0.1% CHAPS, 1% glycerol) was diluted in Assembly Buffer (25 mM HEPES, pH 7.4, 150 mM KCl) to achieve a final protein concentration of 1 mM protein and final KCl concentration of 250mM. Samples were incubated at room temperature for 2 hr to allow formation of oligomeric assemblies. Final concentrations of 8 mM DMTMM (Sigma-Aldrich) were added to the protein samples and incubated at 23 C with shaking for 45 min. Reactions were quenched with 50 mM ammonium bicarbonate and incubated with shaking at room temperature for 15 min. [doi:10.25345/C56Q1ST65] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Centrosomes ; CDK5RAP2 ; SPD-5 ; Polo kinase 1 ; Multivalency ; Phase Separation ; Cross-linking Mass-spectrometry ; in vitro Reconstitution ; Cell Division

Contact

Principal Investigators:
(in alphabetical order) 		Jeffrey Woodruff, UT Southwestern Medical Center, USA
Submitting User: alemoff
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