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Interactome of ANKRD55, an autoimmune disease risk protein with unknown function

frontiersin. 2019; 
Nerea Ugidos, †, Jorge Mena, †, Sara Baquero, Iraide Alloza, Mikel Azkargorta, Felix Elortza and Koen Vandenbroeck, *
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Proteins, Expression, Isolation and Analysis  The supernatant was carefully removed, and nuclei were lysed with RIPA buffer. For the IP of ANKRD55 isoforms together with their binding partners, ANKRD55 and control samples were incubated with Anti-DDK G1 mouse monoclonal antibody coupled to the resin (cat.no. L00432; Genscript) at 4°C overnight (o/n), on rotating wheel. After incubation, resin was washed six times with washing buffer containing 50 mM Tris-HCl and 150 mM NaCl, pH adjusted to 7.4 (phosphatase inhibitors were included for interactome analysis) and ANKRD55 001 complexes were eluted using acidic elution buffer (0.1 M glycine, 0.15 M NaCl and 0,5% SDS pH 2). Get A Quote

摘要

The ankyrin repeat domain-55 (ANKRD55) gene contains intronic single nucleotide polymorphisms (SNPs) associated with risk to contract multiple sclerosis, rheumatoid arthritis or other autoimmune disorders. Risk alleles of these SNPs are associated with higher levels of ANKRD55 in CD4+ T cells. The biological function of ANKRD55 is unknown, but given that ankyrin repeat domains constitute one of the most common protein-protein interaction platforms in nature, it is likely to function in complex with other proteins. Thus, identification of its protein interactomes may provide clues. We identified ANKRD55 interactomes via recombinant overexpression in HEK293 or HeLa cells and mass spectrometry. One hundred forty-... More

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