Signaling pathways altered at the mitotic spindle in clathrin depleted cells

ABSTRACT: We present a snapshot of the regulation of mitotic-spindle proteins and their phosphorylation status in the absence of clathrin during metaphase. A reference list of proteins and phosphosites identified from a mitotic spindle preparation was generated using a label-free and a parallel chemical dimethyl isotopic peptide labelling approach. We further designed experiments to analyse proteins and phosphosites at the mitotic spindle whose abundance was affected by the knock-down of clathrin. We used dimethylation labelling of peptides in combination with fractionation strategies to identify and quantify phosphopeptides enriched by TiO2 beads and the flow-through non-phoshopeptides by ESI-LC-MS/MS for a broad coverage of the mitotic spindle proteome in the absence of clathrin.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Cancer

SUBMITTER: Sushma Rao  

LAB HEAD: A/Prof. Megan Chircop

PROVIDER: PXD001603 | Pride | 2016-05-17

REPOSITORIES: Pride