Past exam of the biochemistry course of the University of Cambridge / 2025 / Paper 5 / Section 1 / 1 / g / Solution

Use a membrane topology measurement on v-SNARE vesicles alone, so that partner binding cannot mask an outward-facing domain. A membrane-impermeant probe directed against the binding-domain region would label only the exposed population while vesicles remain intact. Compare that signal with total accessible protein after lipid bilayer disruption, using a trapped-lumen marker to check that the intact vesicles exclude the probe.

With comparable detection efficiencies and background subtraction,An analogous protease protection assay compares external accessibility with total accessibility. In either approach, lipid bilayer leakage and steric masking must be distinguished from genuinely inward-facing protein.