Project description:KKT23 is a kinetoplastid kinetochore protein that has structural similarity to the GCN5 acetyltransferase domain. We performed crosslinking mass spectrometry of the recombinant KKT3-KKT22-KKT23 complex purified from Sf9 insect cells.
Project description:Settlement-inducing protein complex (SIPC) is a protein that acts as a phoromone cue to attract conspecific of the barnacle Amphibalanus amphitrite to settle on a substrate. Recombinant settlement-inducing protein complex was expressed in insect Sf9 cells through a baculovirus overexpression system, purified and analysed by mass spectrometry to confirm its identity.
Project description:A complex of MAST1 and 14-3-3η was co-expressed and co-purified from Sf9 insect cells. Since the interaction between MAST1 and 14-3-3 is dependent on phosphorylation of MAST1, the complex was analyzed by tandem mass spectrometry to determine which MAST1 residues are phosphorylated.
Project description:Analysis of recombinant mouse Ric-8B protein phosphorylation. Ric-8B was purified from insect cells and E.coli and both treated with protein kinase CK2 and re-purified prior to LC-MS/MS. Another Ric-8B sample from insect cells was not treated with CK2 and also subjected to LC-MS/MS.
Project description:The Nucleosome Remodeling and Deacetylase (NuRD) complex is essential for development in complex animals but has been refractory to detailed analysis because of its low abundance and resistance to recombinant production. In combination with other techniques, crosslinking mass spectrometry was used to elucidate the structure of the Nucleosome Remodelling and Deacetylase (NuRD) complex. Natively-purified NuRD, and four recombinantly-expressed NuRD subcomplexes, and a PWWP2A-MTA-HDAC-RBBP alternative ‘NuRD-like’ complex were subjected to crosslinking with DSS, ADH, BS3 and DMTMM.
Project description:We performed chemical crosslinking mass spectrometry of native protein complexes purified from Trypanosoma brucei procyclic cells to obtain protein-protein interaction information for the chromosomal passenger complex (CPC) and kinetochore complex. Crosslinks among CPC subunits were detected using YFP-AUK1, while those among KKT8 complex subunits were detected in the YFP-KKIP1 sample. We also performed immunoprecipitation and mass spectrometry of CPC subunits (full-length proteins and fragments) to identify co-purifying proteins.