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. 2004 Feb;78(3):1109–1120. doi: 10.1128/JVI.78.3.1109-1120.2004

FIG. 4.

FIG. 4.

MARCH proteins act as ubiquitin ligases in vitro. GST fusion proteins of the amino-terminal domains of individual MARCH proteins were incubated with ubiquitin, ATP, E1, and the indicated ubiquitin-conjugating enzymes (ubc, E2) and separated by SDS-polyacrylamide gel electrophoresis prior to immunoblotting with the ubiquitin-specific antibody P4D1. Ubiquitin ligase activity is indicated by the appearance of high-molecular-weight ubiquitinated species. Ubiquitinated proteins were not detected in the absence of E3s (None) or with GST alone (unpublished data).

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