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Duck RIG-I CARD domains are ubiquitinated at K167 and K193.

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posted on 2014-01-23, 04:42 authored by Domingo Miranzo-Navarro, Katharine E. Magor

A. Coomassie stained gel showing the different ubiquitinated forms of the duck RIG-I CARD domains. B. MaxEnt3 deconvoluted MS/MS spectra of a peptide bearing the typical diglycine signal for ubiquitination at Lys 193, showing the singly-charged forms of the B and Y ions detected. C. Three-dimensional detail of duck RIG-I structure in the region of Q170 (red), corresponding to human K172, and the closest lysine, K167 (blue) (PDB code:4a2w) [31]. Figure was created using Protean 3D from DNA Lasergene 9. D. MaxEnt3 deconvoluted MS/MS spectra of a peptide showing the ubiquitination signal for K167. E. GST pulldown of duck CARD domains from cells co-transfected with HA-ubiquitin, or HA-ubiquitin-K0 (all lysines mutated) or HA-ubiquitin K63 (only lysine 63 intact). Band 2 in the GST pulldown is a doublet and the lower band is missing in the absence of K63-linked ubiquitin chains. F. MaxEnt3 deconvoluted MS/MS spectra of a peptide of ubiquitin recovered from band 2 showing the diglycine signal at Lys 63, indicating the presence of Lys 63-linked polyubiquitin.

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