Skip to main content
Fig. 5 | BMC Biochemistry

Fig. 5

From: Kv1.3 contains an alternative C-terminal ER exit motif and is recruited into COPII vesicles by Sec24a

Fig. 5

In vitro Kv1.3-Sec24a membrane floatation assay. Membrane floatation assay used to test for the association between Kv1.3 and Sec24a341. (a) Kv1.3 proteins reconstituted into synthetic lipid vesicles (proteoliposomes) and (b) control lipid vesicles (liposomes). (c) Schematic of the floatation assay. Proteoliposomes, drawn as small black circles, migrate through the three-step sucrose gradient (0 %, 25 %, and 30 % w/v sucrose; top (1), middle (2) and bottom (3), respectively) after incubation and centrifugation. (d) Kv1.3 proteoliposomes were found in the top fraction after centrifugation. (e) When Kv1.3 proteoliposomes (~65 kDa as a monomer) were incubated with Sec24a341 (~80 kDa), both Kv1.3 and Sec24a341 were detected in the top fraction. (f) Sec24a341 alone was not detected in the top fraction. (g) When Kv1.3 proteins in detergent micelles were mixed with Sec24a341 in the presence of control liposomes, both Kv1.3 and Sec24a341 were found in the top fraction. (h) Sec24a341 incubated with control liposomes was not found in the top fraction. (i) Kv1.3 micelles and Sec24a341 do not float in the absence of membranes. (j) Kv1.3 was detected in the top fraction when Kv1.3 in detergent micelles were incubated with control liposomes. Scale bar = 100 nm

Back to article page