Fig. 5: Effect of Ca2+ on membrane binding of TRIM72.
From: Structure and activation of the RING E3 ubiquitin ligase TRIM72 on the membrane
a, Protein–lipid overlay analysis showing the interference. TRIM72 proteins were incubated with lipid strips in the presence or absence of Ca2+ and/or EGTA and then probed with an anti-TRIM72 antibody. PE, phosphatidylethanolamine. N/A indicates phospholipid is not applied. b, Flow cytometry analysis comparing the effects of Ca2+ on TRIM72 and annexin V (ANXV). PS liposomes were preloaded with Ca2+ or loaded after Ca2+ exposure and then incubated with TRIM72 or annexin V. Binding was detected using fluorescently labeled sfGFP–TRIM72 and fluorescein isothiocyanate (FITC)–annexin V. Ex488/Em533 nm, excitation at 488 nm/emission at 533 nm. c, Quantification of the data in b. Experiments were performed in triplicate. Data are presented as mean values ± s.d. The PS concentration was 30 mol% in PS liposomes. Ca2+ ion, yellow; TRIM72, green; annexin V, pink.
