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A central helical hairpin in SPD-5 enables centrosome strength and assembly

ABSTRACT: Cross-linked samples consist of middle region of SPD-5 (541-677) plus 18 amino acids at the N-terminus (MGSSHHHHHHENLYFQSN) which we term F1. Two F1 samples (7.6 uM) were incubated with either kinase dead or constitutively active PLK-1 plus ATP-MgCl2 in a 150 mM KCl, 25mM HEPES, pH7.4 buffer for 2 hours at room temperature. Samples were then cross-linked using 8mM DMTMM for 45min and quenched with 50 mM ammonium bicarbonate for 15 min at RT shaking at 300rpm. Samples ran on a 16-20% SDS-page gel revealing a monomer band and a dimer band in both conditions using a coomassie based stain. Monomer bands (LUM2_1076612, PLK-1 KD, LUM2_1076614, PLK-1 -CA) and dimer bands (LUM2_1076613, PLK-1 KD, LUM2_1076615, PLK-1 KD) were excised.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Caenorhabditis Elegans (ncbitaxon:6239)

SUBMITTER: Jeffrey Woodruff

PROVIDER: MSV000094191 | MassIVE | Wed Feb 28 09:57:00 GMT 2024

REPOSITORIES: MassIVE

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