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. Author manuscript; available in PMC: 2019 Jun 20.
Published in final edited form as: Annu Rev Biochem. 2017 Aug 25;87:783–807. doi: 10.1146/annurev-biochem-062917-011852

Figure 9.

Figure 9

Tryptic digestion of sealed membrane vesicles from cells expressing wild-type (WT) Scap or two mutants (Y234A in Loop 1 and Y640S in Loop 7) that prevent interloop binding. Scap-deficient CHO cells transfected with the indicated Scap plasmid were depleted of sterols and incubated for 4 h without or with 50 μM cholesterol complexed to methyl-β-cyclodextrin (MCD). Sealed membrane vesicles were prepared and treated with trypsin. Samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted with either (a) anti-Loop 1 antibody or (b) anti-Loop 7 antibody. The L4 band in panel a denotes trypsin cleavage in Loop 4. The R505 band in panel b denotes cleavage at arginine-505 in Loop 6. Reprinted from Reference 63.