TY - GEN
T1 - Monitoring receptor trafficking following retromer and WASH deregulation
AU - Osborne, Douglas G.
AU - Phillips-Krawczak, Christine A.
AU - Billadeau, Daniel D.
PY - 2015
Y1 - 2015
N2 - Cell surface receptors that have been internalized and enter the endocytic pathway have multiple fates including entrance into the multivesicular body pathway on their way to lysosomal degradation, recycling back to the cell surface, or retrograde trafficking out of the endolysosomal system back to the Golgi apparatus. Two ubiquitously expressed protein complexes, WASH and the endosomal coat complex retromer, function together to play a central role in directing the fate of receptors into the latter two pathways. In this chapter, we describe fluorescent- and flow cytometry-based methods for analyzing the recycling and retrograde trafficking of two receptors, α5β1 and CI-M6PR, whose intracellular fates are regulated by WASH and retromer activity. The guidelines presented in this chapter can be applied to the analysis of any cell surface or intracellular membrane protein to determine the impact of WASH or retromer deregulation on its intracellular trafficking route.
AB - Cell surface receptors that have been internalized and enter the endocytic pathway have multiple fates including entrance into the multivesicular body pathway on their way to lysosomal degradation, recycling back to the cell surface, or retrograde trafficking out of the endolysosomal system back to the Golgi apparatus. Two ubiquitously expressed protein complexes, WASH and the endosomal coat complex retromer, function together to play a central role in directing the fate of receptors into the latter two pathways. In this chapter, we describe fluorescent- and flow cytometry-based methods for analyzing the recycling and retrograde trafficking of two receptors, α5β1 and CI-M6PR, whose intracellular fates are regulated by WASH and retromer activity. The guidelines presented in this chapter can be applied to the analysis of any cell surface or intracellular membrane protein to determine the impact of WASH or retromer deregulation on its intracellular trafficking route.
KW - Endosome
KW - F-actin
KW - Receptor trafficking
KW - Retromer
KW - WASH
UR - http://www.scopus.com/inward/record.url?scp=84945451765&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84945451765&partnerID=8YFLogxK
U2 - 10.1016/bs.mcb.2015.03.018
DO - 10.1016/bs.mcb.2015.03.018
M3 - Conference contribution
C2 - 26360036
AN - SCOPUS:84945451765
SN - 9780128028292
T3 - Methods in Cell Biology
SP - 199
EP - 213
BT - Methods in Cell Biology
A2 - Guo, Wei
PB - Academic Press Inc
ER -