PMID- 22325598
OWN - NLM
STAT- MEDLINE
DCOM- 20120531
LR  - 20150813
IS  - 0091-679X (Print)
IS  - 0091-679X (Linking)
VI  - 108
DP  - 2012
TI  - Reconstituting multivesicular body biogenesis with purified components.
PG  - 73-92
LID - 10.1016/B978-0-12-386487-1.00004-3 [doi]
AB  - Activated cell surface receptors are rapidly removed from the plasma membrane 
      through clathrin mediated endocytosis and transported to the endosome where they 
      are either recycled or sorted to the lysosomal pathway to be degraded. Receptors, 
      destined for degradation in the lysosome, are packaged into intraluminal vesicles 
      (ILVs) of endosomes by a reaction that is topologically unrelated to other 
      budding reactions in cells. First, receptors are clustered at the endosomal 
      membrane and receptor-rich membrane patches then bud towards the lumen of the 
      endosome. The nascent membrane buds are finally cleaved from the limiting 
      membrane to release cargo-bearing vesicles into the endosomal interior. The 
      molecular machinery that drives multivesicular body biogenesis, the endosomal 
      sorting complex required for transport (ESCRT) machinery, has been identified 
      through genetic screens. It consists of the cytoplasmic, hetero-multimeric 
      complexes ESCRT-0, -I, -II, and -III, and of the Vps4/VtaI complex. Although the 
      ESCRT machinery has been characterized extensively using cell-biological and 
      biochemical approaches, the molecular mechanism of multivesicular body biogenesis 
      remained unclear. In this chapter, I will present in vitro reconstitution systems 
      that we used to study ESCRT-driven membrane remodeling reactions with purified 
      components on artificial membranes. This includes generation of large and giant 
      unilamellar liposomes, as well as in vitro reconstitution reactions of 
      fluorescently labeled proteins on such membranes. I will discuss both, the 
      potential of in vitro systems to analyze membrane-remodeling events and also 
      their limitations.
CI  - Copyright (c) 2012 Elsevier Inc. All rights reserved.
FAU - Wollert, Thomas
AU  - Wollert T
AD  - Molecular Membrane and Organelle Biology, Max Planck Institute of Biochemistry, 
      Martinsried, Germany.
LA  - eng
GR  - Intramural NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Intramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Cell Biol
JT  - Methods in cell biology
JID - 0373334
RN  - 0 (Endosomal Sorting Complexes Required for Transport)
RN  - 0 (Membrane Lipids)
RN  - 0 (Membrane Proteins)
RN  - 0 (Unilamellar Liposomes)
SB  - IM
MH  - Algorithms
MH  - Animals
MH  - Biological Transport
MH  - Endosomal Sorting Complexes Required for Transport/*chemistry
MH  - Fluorescence Recovery After Photobleaching
MH  - Humans
MH  - Membrane Lipids/chemistry
MH  - Membrane Proteins/chemistry
MH  - Models, Biological
MH  - Multivesicular Bodies/*chemistry
MH  - Protein Binding
MH  - Unilamellar Liposomes/chemistry
EDAT- 2012/02/14 06:00
MHDA- 2012/06/01 06:00
CRDT- 2012/02/14 06:00
PHST- 2012/02/14 06:00 [entrez]
PHST- 2012/02/14 06:00 [pubmed]
PHST- 2012/06/01 06:00 [medline]
AID - B9780123864871000043 [pii]
AID - 10.1016/B978-0-12-386487-1.00004-3 [doi]
PST - ppublish
SO  - Methods Cell Biol. 2012;108:73-92. doi: 10.1016/B978-0-12-386487-1.00004-3.