Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

HEK cell MYO7A-PDZD7 immunoprecipitation

ABSTRACT: Through its activity in stereocilia, MYO7A (myosin VIIA) is essential for hair cell function in the inner ear. Utilizing multiple stages of immunoaffinity enrichment, we have developed a strategy that allows us to partially purify stereocilia membranes from thousands of chick inner ears and isolate low-abundance MYO7A protein complexes from those membranes, including one with the scaffolding protein PDZD7. To confirm the interaction of PDZD7 and MYO7A, we expressed them singly or together in HEK cells, and immunoprecipitated detergent-soluble fractions with specific antibodies. PDZD7 and MYO7A interacted strongly in these tissue-culture cells.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Permanent Cell Line Cell, Early Embryonic Cell

SUBMITTER: Peter Barr-Gillespie

LAB HEAD: Peter Barr-Gillespie

PROVIDER: PXD004266 | Pride | 2016-08-16

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	GILL-102_Sample7_HEKMYO7aGFPTOTAL_Shotgun_Run1_25cm_120min_20150814_OT.raw	Raw
	GILL-102_Sample7_HEKMYO7aGFPTOTAL_Shotgun_Run2_25cm_120min_20150814_OT.raw	Raw
	GILL-102_Sample8_HEKMYO7aGFPCONIP_Shotgun_Run1_25cm_120min_20150814_OT.raw	Raw
	GILL-102_Sample8_HEKMYO7aGFPCONIP_Shotgun_Run2_25cm_120min_20150814_OT.raw	Raw
	GILL-102_Sample9_HEKMYO7aGFPMYO7aIP_Shotgun_Run1_25cm_120min_20150814_OT.raw	Raw

Items per page:

1 - 5 of 32

Similar Datasets

D10 Stereocilia Membrane Enrichment

Project description:Through its activity in stereocilia, MYO7A (myosin VIIA) is essential for hair cell function in the inner ear. Utilizing multiple stages of immunoaffinity enrichment, we have developed a strategy that allows us to partially purify stereocilia membranes from thousands of chick inner ears and isolate low-abundance MYO7A protein complexes from those membranes. The D10 stereocilia membrane enrichment protocol involves density centrifugation steps and immuno-enrichment of stereocilia using the D10 antibody, directed against the major stereocilia transmembrane protein PTPRQ. The data in this submission document the effectiveness of the enrichment procedure.

2016-08-16 | PXD004222 | Pride

Myosin VIIA 138-1 immunoaffinity purification

Project description:Through its activity in stereocilia, MYO7A (myosin VIIA) is essential for hair cell function in the inner ear. Utilizing multiple stages of immunoaffinity enrichment, we have developed a strategy that allows us to partially purify stereocilia membranes from thousands of chick inner ears and isolate low-abundance MYO7A protein complexes from those membranes. By analysis of MYO7A and co-purifying molecules with shotgun and targeted mass spectrometry, we demonstrated that MYO7A forms a complex with PDZD7, a paralog of USH1C and DFNB31, which has been shown to localize to the ankle-link region of stereocilia.

2016-08-16 | PXD004221 | Pride

Mouse Stereocilia Capping Protein Immunoaffinity Purification

Project description:Capping protein controls stereocilia length and width during hair bundle development. To determine what other proteins are involved in capping protein regulation, we carried out immunoaffinity purifications targeted at either CAPZA or CAPZB2. The starting material for immunopurification was crude stereocilia membranes isolated from mouse inner ear.

2017-08-25 | PXD006077 | Pride

GIPC3 AP-MS/MS from chick inner ear extracts

Project description:To understand how GIPC3 exerts its effects on cuticular plate dimensions, we examined the GIPC3 protein-interaction network in hair cells. We immunoaffinity purified GIPC3 from crosslinked chicken inner ear extracts that were enriched for stereocilia, but still contain large amounts of hair-cell cytoplasmic proteins (Morgan et al., 2016). We carried out two separate experiments, each with ~1000 chicken ears, where we stabilized protein complexes using primary amine-reactive homo-bifunctional N-hydroxysuccimide ester crosslinkers that are thiol-cleavable and hence reversible (Mattson et al., 1993). In one experiment, we used dithiobis(succinimidyl propionate) (DSP), a membrane-permeable crosslinker that crosslinks extracellular and intracellular complexes; in the other experiment, we used 3,3'-dithiobis(sulfosuccinimidyl propionate) (DTSSP), which is membrane impermeant and thus only stabilizes extracellular and transmembrane complexes. We prepared soluble extracts of crude, crosslinked stereocilia and used these fractions for identifying GIPC3-interacting proteins.

2023-05-09 | PXD038234 | Pride

Chick utricle hair bundles and utricular epithelium—Velos

Project description:Purified hair bundles and utricular epithelium from E19-E20 chick inner ears were analyzed by LC-MS/MS to determine the abundant and enriched proteins of the hair bundle. This dataset used a Thermo LTQ Velos mass spectrometer for protein detection.

2015-11-17 | PXD002414 | Pride

Chick utricle hair bundles and utricular epithelium—LTQ

2015-11-17 | PXD002410 | Pride

Reprocessed: Chick utricle hair bundles and utricular epithelium—Orbitrap

Project description:This is a reanalysis of PXD000104. Purified hair bundles and utricular epithelium from E19-E20 chick inner ears were analyzed by LC-MS/MS to determine the abundant and enriched proteins of the hair bundle. This dataset used a Thermo Orbitrap Elite mass spectrometer for protein detection.

2015-11-17 | PXD002445 | Pride

Transcription profiling of inner ears from Errb mutant mice and wild type littermate controls

Project description:Transcriptional profiles were compared in microdissected lateral walls of the inner ears from Errb mutant mice and wild type littermate controls. The goal is to identify transcriptional targets of Errb and candidate genes for inner ear diseases. Experiment Overall Design: Errb mutant mice were generated by conditional knock-out strategy. Inner ears from 10 mice were pooled for each sample and 3 replicates of wild type and mutant samples were analyzed.

2008-06-16 | E-GEOD-8434 | biostudies-arrayexpress

Wild-type and COUP-TFI-/- newborn inner ear microarrays

Project description:In order to establish a list of candidate direct COUP-TFI gene targets in the inner ear, we analyzed the differential gene expression profiles of the wild-type and the COUP-TFI–/– P0 inner ears.

2010-05-31 | GSE16744 | GEO

Transcriptional regulation by Errb (Nr3b2) in stria vascularis

2007-08-01 | GSE8434 | GEO