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Thyroglobulin interactome profiling defines altered proteostasis topology associated with thyroid dyshormonogenesis

Mol Cell Proteomics. 2020-11; 
Madison T Wright, Logan Kouba, Lars Plate
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Plasmid DNA Preparation … EXPERIMENTAL PROCEDURES Plasmids and Antibodies FLAG-tagged (FT)-Tg in pcDNA3.1+/C-(K)-DYK plasmid was purchased from Genscript (Clone ID OHu20241) … (Sigma, A2220) or G1 Anti-DYKDDDDK affinity resin (GenScript, L00432) overnight at 4°C while rocking … Get A Quote

摘要

Thyroglobulin (Tg) is a secreted iodoglycoprotein serving as the precursor for T3 and T4 hormones. Many characterized Tg gene mutations produce secretion-defective variants resulting in congenital hypothyroidism (CH). Tg processing and secretion is controlled by extensive interactions with chaperone, trafficking, and degradation factors comprising the secretory proteostasis network. While dependencies on individual proteostasis network components are known, the integration of proteostasis pathways mediating Tg protein quality control and the molecular basis of mutant Tg misprocessing remain poorly understood. We employ a multiplexed quantitative affinity purification-mass spectrometry approach to define the Tg ... More

关键词

Affinity proteomics, Cell secretion*, Congenital Hypothyrodism, Interactomics, Protein Folding*, Protein-Protein Interactions*, Proteostasis, Tandem Mass Spectrometry, Tandem Mass Tags