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Algae metagenome

ABSTRACT: Pleurocapsa minor GSE-CHR-MK-17-07R and co-occurring heterotrophs from sandstone from Lower Calf Creek Falls, Grand Staircase Escalante National Monument, UT, United States - 20190625_44

PROVIDER: PRJNA677054 | ENA |

REPOSITORIES: ENA

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			Action	DRS
		Other
	SRR13207074_1.fastq.gz	Fastqsanger.gz
	SRR13207074_2.fastq.gz	Fastqsanger.gz

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Identification of the UT-A1 Urea Channel Interactome

Project description:The urea channel Slc14a2 (or UT-A1) mediates vasopressin-regulated urea transport across the inner medullary collecting duct (IMCD). Previously, UT-A1 was found to present in a high molecular weight complex, suggesting UT-A1 is involved in certain protein-protein interactions. The present study sought to identify the proteins that interact with UT-A1 in this complex for a better understanding of how UT-A1 is regulated. Rat IMCD suspensions were treated with or without V2 receptor agonist, dDAVP, followed by in-cell crosslinking using BSOCOES and detergent solubilization. Immunoprecipitation using Dynabeads coated with UT-A1 specific antibody successfully pulled down the UT-A1 proteins. In-gel digestion protocol was carried out to prepare samples for liquid chromatographic mass spectrometry analysis of tryptic peptides using a Velos-Orbitrap mass spectrometer. The peptides passing stringent spectral quality thresholds were quantified (label-free) to identify those with (UTA-1 antibody/preimmune IgG) >4. A total of 128 UT-A1 interacting proteins were identified. Gene Ontology analysis maps the distribution of these proteins throughout major cell compartments: endoplasmic reticulum, Golgi, endosomes, cytosol and plasma membrane. Among them are four protein kinases (Cdc42bpb, Phkb, Camk2d, Mtor) that play roles in vasopressin-regulated phosphorylation of UT-A1. Non-label quantification was also performed to determine the stoichiometry of UT-A3 with UT-A1, the result does not support an oligomeric complex formation of UT-A1/A3. In conclusion, we have provided a refined list of UT-A1 binding proteins which can be useful for further analysis of the vasopressin signaling pathway in regulation of UT-A1 in IMCD.

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Project description:We sampled lake-type and riverine sockeye in the pristine natural habitats of Aniakchak National Monument and Preserve, Alaska USA.

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