Searching for binding partners of the SLX4 577-795 region and assessing the impact of the SLX4 D614G and L618P cancer patient-derived mutations on their association with SLX4

ABSTRACT: The project was aimed at identifying new binding partners of SLX4 that associate with a region spanning the conserved amphipathic helix of SLX4 and the downstream BTB domain. It was also aimed at assessing which of those partners might have their association with SLX4 abrogated by the cancer patient-derived D614G and L618P SLX4 mutations. The goal was ultimately to determine whether RTEL1 which we had identified as a novel binding partner of SLX4 and that does not bind SLX4 D614G or SLX4 L618P mutant proteins, is the only SLX4 binding partner to be impacted by those mutations. A YFP-SLX4 577-795 wt or mutated fragment produced in Hela Flp-In TRex cells was immunoprecipitated using a GFP-nanobody. YFP-pull downs were washed 5 times with 50 mM Tris-HCl [pH 8.0] buffer elution of the proteins bound to the GFP-nanobody directly in NuPAGE LDS sample buffer (Invitrogen) for 5 min at 95˚C.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Luc Camoin  

LAB HEAD: MarseilleProteomics CRCM and Gaillard PH Lab

PROVIDER: PXD012426 | Pride | 2020-03-05

REPOSITORIES: Pride