Proteomics

Dataset Information

0

MudPIT analyses of the human proteins biotinylated by proximity to dynein/dynactin subunits fused to biotin ligase (BioID)

ABSTRACT: We identified the human dynein proteome by attaching a promiscuous biotin ligase (BioID) to 7 distinct components of the dynein machinery, including a subunit of its essential cofactor dynactin. BioID experiments were performed with stable HEK293 cell lines expressing dynein (IC1, IC2, LIC1, LIC2, RB, TcTex-1) and dynactin (p62) subunits tagged with BioID-3X FLAG. BioID-only transfected cells were used as negative controls. For BioID experiments, cells were lysed in the presence of detergents to disrupt the dynein/dynactin complex, allowing the identification of proteins that were proximal to the tagged subunit prior to cell lysis. After purification of biotinylated proteins on streptavidin-conjugated beads, the eluates were precipitated with TCA. After washing with acetone, the protein mixtures were digested with endoproteinase Lys-C and trypsin (Roche) and analyzed by MudPIT. We performed BioID purification followed by MudPIT in quadruplicate and used a label-free quantitative proteomics approach to calculate the enrichment of each identified protein relative to BioID control replicates.

INSTRUMENT(S): LTQ

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Laurence Florens  

PROVIDER: MSV000080759 | MassIVE | Wed Mar 29 09:59:00 BST 2017

SECONDARY ACCESSION(S): PXD006206

REPOSITORIES: MassIVE

Json Xml

Dataset's files

Source:
Action DRS
Other
Items per page:
1 - 1 of 1
altmetric image

Publications

In human cells, cytoplasmic dynein-1 is essential for long-distance transport of many cargos, including organelles, RNAs, proteins, and viruses, towards microtubule minus ends. To understand how a single motor achieves cargo specificity, we identified the human dynein interactome by attaching a promiscuous biotin ligase ('BioID') to seven components of the dynein machinery, including a subunit of the essential cofactor dynactin. This method reported spatial information about the large cytosolic  ...[more]

Similar Datasets

Proteomics MudPIT analyses of the human proteins biotinylated by proximity to biotin ligase (BioID) fused to the N-terminus of activators of the dynein/dynactin motor
2017-03-30 | MSV000080807 | MassIVE
Proteomics MudPIT analyses of the human proteins biotinylated by proximity to biotin ligase (BioID) fused to the C-terminus of activators of the dynein/dynactin motor
2017-03-30 | MSV000080808 | MassIVE
Transcriptomics Specific Roles for the Ccr4-Not Complex Subunits in Expression of the Genome
2008-12-10 | GSE10521 | GEO
Genomics Genome-wide localization of the RNA polymerase III transcription machinery in human cells
2010-02-11 | GSE18184 | GEO
Proteomics Side-by-side comparison of AP-MS, Tri-functional affinity probe, BioID, PUP-IT and SPIDER to identify CheZ-interacting proteins
2022-11-03 | PXD037952 |
Transcriptomics Specific Roles for the Ccr4-Not Complex Subunits in Expression of the Genome
2008-12-10 | E-GEOD-10521 | biostudies-arrayexpress
Transcriptomics ActD timecourse in HeLa cells, HeLa RNF219 KO cells, HeLa RNF219 KO cells overexpressing 3xFLAG-SBP-RNF219-WT or 3xFLAG-SBP-RNF219-mut
2021-10-27 | GSE172019 | GEO
Transcriptomics Profiling the endothelial translatome in vivo using ‘AngioTag’ zebrafish
2019-03-29 | GSE121973 | GEO
Proteomics Human Ku70 WT AP-MS - Mapping the Ku interactome using proximity-dependent biotin identification in human cells
2019-01-03 | PXD010930 | Pride
Proteomics MudPIT analyses of the human proteins biotinylated by proximity to exogenously expressed biotin ligase (BioID)
2017-03-30 | MSV000080809 | MassIVE