Pregled bibliografske jedinice broj: 589006
Examining Protein Interactions Using the Large- Scale Pull-down Approach and Endogenously Tagged Yeast Strains
Examining Protein Interactions Using the Large- Scale Pull-down Approach and Endogenously Tagged Yeast Strains // 2006 Yeast Genetics and Molecular Biology Meeting
Princeton (NJ), Sjedinjene Američke Države, 2006. (poster, međunarodna recenzija, sažetak, znanstveni)
CROSBI ID: 589006 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Examining Protein Interactions Using the Large- Scale Pull-down Approach and Endogenously Tagged Yeast Strains
Autori
Suter, Bernhard ; Fetchko, Michael ; Imhof, Ralph ; Stoffel-Studer Ingrid ; Zbinden, Caroline ; Beneti, Lucija ; Hort, Jacqueline ; Stagljar, Igor
Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni
Skup
2006 Yeast Genetics and Molecular Biology Meeting
Mjesto i datum
Princeton (NJ), Sjedinjene Američke Države, 25.07.2006. - 30.07.2006
Vrsta sudjelovanja
Poster
Vrsta recenzije
Međunarodna recenzija
Ključne riječi
protein-protein interactions; cross and capture system
Sažetak
The organization of the cellular proteome into multiprotein complexes is a major focus for functional proteomics. In yeast Saccharomyces cerevisae, systematic approaches have proven to be successful to determine large protein assemblies and to assign new functions to the associated proteins. Here, we present a novel approach (“Cross and Capture”) that allows a rapid assessment of protein-protein interactions in yeast using two sets of differently tagged yeast arrays in the two haploid mating types. Open reading frames (ORFs) in the a-haplotype (bait) array were tagged with six consecutive histidines (6xHIS), whereas ORFs in the ?-haplotype (prey) array were tagged with the 3xVSV epitope, respectively. In order to determine protein- protein interactions, a specific bait strain is selected and mated with a set of strains containing potential preys. Protein extracts from the diploids containing both tags are then incubated with nickel beads that bind to the 6xHIS tag and thus allow the“pull-down” of bait and bait-associated proteins. Association of the selected prey proteins is then verified by probing Western Blots with anti-VSV antibody. So far we have generated and verified two sets of 500 bait and prey strains that comprise known DNA replication, repair and recombination proteins, but also a large number of yeast nuclear ORFs of unascribed function. “Cross and Capture” was successfully used to detect a number of previously described multiprotein complexes and extensive screenings for novel protein-protein interactions are currently underway.
Izvorni jezik
Engleski
Znanstvena područja
Biologija
