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Highly efficient overexpression and purification of multisubunit tethering complexes in Saccharomyces cerevisiae.
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Article Synopsis
- Vesicle trafficking is a crucial process in eukaryotic cells, involving the exchange of materials between organelles, and multisubunit tethering complexes (MTCs), like the HOPS complex, play a key role in this.
- The study developed an efficient method to overexpress and purify the HOPS complex from yeast Saccharomyces cerevisiae by integrating a strong promoter before each subunit and using advanced purification techniques.
- The researchers achieved significantly higher yields of the HOPS complex compared to previous methods and also successfully purified two other MTCs, paving the way for more detailed biochemical and functional studies in the future.
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Numerous mutations that impair retrograde membrane trafficking between endosomes and the Golgi apparatus lead to neurodegenerative diseases. For example, mutations in the endosomal retromer complex are implicated in Alzheimer's and Parkinson's diseases, and mutations of the Golgi-associated retrograde protein (GARP) complex cause progressive cerebello-cerebral atrophy type 2 (PCCA2). However, how these mutations cause neurodegeneration is unknown.
View Article and Find Full Text PDFAre all multisubunit tethering complexes bona fide tethers?
Traffic
November 2014
Department of Biology, Concordia University, Montreal, Quebec, Canada.
Since the late 1990s, a number of multisubunit tethering complexes (MTCs) have been described that function in membrane trafficking events: TRAPP I, TRAPP II, TRAPP III, COG, HOPS, CORVET, Dsl1, GARP and exocyst. On the basis of structural and sequence similarities, they have been categorized as complexes associated with tethering containing helical rods (CATCHR) (Dsl1, COG, GARP and exocyst) or non-CATCHR (TRAPP I, II and III, HOPS and CORVET) complexes (Yu IM, Hughson FM. Tethering factors as organizers of intracellular vesicular traffic.
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