A secretion system for cargo protein identification of vacuolar sorting receptors

Abstract

Vacuolar sorting receptors (VSRs) are type I integral membrane family proteins in plant cells that can sort cargo proteins at the late Golgi or trans-Golgi network (TGN) for vacuolar transport via the prevacuolar compartment (PVC). However, little is known about VSR cargo proteins in plants. Here, we describe a new method for the identification of VSR cargos, which is based on the premise that the expressed N-terminus of VSRs will be secreted into the culture media along with their corresponding cargo proteins. The protocol described here should be applicable to all VSRs and should be also useful for other receptor cargo identification and protein-protein interaction in vivo.