BAIT

SIR3

CMT1, MAR2, STE8, chromatin-silencing protein SIR3, L000001896, YLR442C
Silencing protein; interacts with Sir2p, Sir4p, and histone H3 and H4 tails to establish transcriptionally silent chromatin state; required for spreading of silenced chromatin; recruited to chromatin through interaction with Rap1p; C-terminus (residues 840-978) assumes variant winged helix-turn-helix (wH) fold that mediates homodimerization, which is critical for holo-SIR complex loading; SIR3 has a paralog, ORC1, that arose from the whole genome duplication
Saccharomyces cerevisiae (S288c)
PREY

ACO1

GLU1, aconitate hydratase ACO1, L000000022, YLR304C
Aconitase; required for the tricarboxylic acid (TCA) cycle and also independently required for mitochondrial genome maintenance; phosphorylated; component of the mitochondrial nucleoid; mutation leads to glutamate auxotrophy
GO Process (2)
GO Function (3)
GO Component (4)
Saccharomyces cerevisiae (S288c)

Cross-Linking-MS (XL-MS)

An interaction is detected between two proteins using chemically reactive or photo-activatable cross-linking reagents that covalently link amino acids in close proximity, followed by mass spectrometry analysis to identify the linked peptides (reviewed in PMID 37406423, 37104977). Experiments may be carried with live cells or cell lysates in which all proteins are expressed at endogenous levels (e.g. PMID 34349018, 35235311) or with recombinant proteins (e.g., PMID 28537071).

Publication

Improving Identification of In-organello Protein-Protein Interactions Using an Affinity-enrichable, Isotopically Coded, and Mass Spectrometry-cleavable Chemical Crosslinker.

Makepeace KAT, Mohammed Y, Rudashevskaya EL, Petrotchenko EV, Voegtle FN, Meisinger C, Sickmann A, Borchers CH

An experimental and computational approach for identification of protein-protein interactions by ex vivo chemical crosslinking and mass spectrometry (CLMS) has been developed that takes advantage of the specific characteristics of cyanurbiotindipropionylsuccinimide (CBDPS), an affinity-tagged isotopically coded mass spectrometry (MS)-cleavable crosslinking reagent. Utilizing this reagent in combination with a crosslinker-specific data-dependent acquisition strategy based on MS2 scans, and a software pipeline ... [more]

Mol Cell Proteomics Apr. 01, 2020; 19(4);624-639 [Pubmed: 32051233]

Throughput

  • High Throughput

Additional Notes

  • authors used a cut-off of FDR 2% to determine hit genes

Curated By

  • BioGRID