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VEX1-GFP Pull-downs - Affinity-purified VEX1GFP-associated proteins using cryomilling and high-affinity nanobodies

ABSTRACT: We affinity-purified VEX1GFP-associated proteins using cryomilling and high-affinity nanobodies. The procedure was carried out initially in insect-stage T. brucei, for which protocols were established, and then in bloodstream-form T. brucei. Quantitative proteomic analysis revealed tag-dependent enrichment of GFP and the same set of five proteins in four independent experiments; VEX1 was enriched, as expected, but also Tb927.11.13380, an ortholog of the nonsense-mediated mRNA-decay (NMD) ATP-dependent superfamily 1-type helicases, UPF1/SMG2/NAM7/Rent1 19), now designated VEX2 (predicted 224-kDa, 2026 residues). The other three proteins that displayed tag-dependent enrichment were all three components of chromatin assembly factor 1 (CAF-1; CAF-1a, Tb927.8.3980; CAF-1b, Tb927.10.7050, CAF-1c, Tb927.11.4970a), the evolutionary conserved hetero-trimeric replication-associated histone chaperone.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Trypanosoma Brucei

DISEASE(S): Trypanosomiasis

SUBMITTER: Joana Faria

LAB HEAD: David Horn

PROVIDER: PXD013304 | Pride | 2019-05-31

REPOSITORIES: Pride

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