Post on 15-Aug-2015
transcript
Cell Biology
S. Rahgozar,PhD
University of Isfahan
Faculty of Science
4. Protein sorting and transport
4.3. Lysosomes and Vesicular transport
1392-93
Vesicular transport plays a central role in the traffic of molecules
Why is it important?
The selectivity of such transport is key to maintaining the functional
organization of the cell
Experimental approaches?
Isolation of yeast mutants defective in protein transport and sorting
Reconstitution of vesicular transport in cell-free systems
Biochemical analysis of synaptic vesicles
Tracing the path of specific GFP fusion proteins through the
secretory network
Proteomic analysis of specific secretory compartments
between membrane-enclosed compartments
from the cell surface
Formation and fusion of a
transport vesicles
Cargo selection,
Coat proteins,
Vesicle budding
The coats
assemble as the secretory
protein-containing vesicle
buds off the donor membrane
are removed from the vesicle
in the cytosol before it reaches
its target.
Three important aspects in cargo selection and transport
Small GTP-binding proteins (transferred from cytosol)
ADP-ribosylation factors
o ARFs 1-3
o Sar1
Rab proteins (for docking of transport vesicles)
Adaptor proteins
Coat proteins
o Clathrin
o COPI
o COPII
(for vesicular budding)
Regulation of monomeric GTPase
RGS
RGS : Regulators of G protein
signaling protein. Also
called GAP (GTPase-
activating proteins)
GEF: Guanine nucleotide
exchange factor
Transport by coated vesicles
COP: coat protein
Golgi apparatus
Golgi
stack
Golgi in a fibroblast
Initiation of a clathrin-coated
vesicle by ARF1
GGA: adaptor protein
AP1: adaptor protein
Incorporation of lysosomal proteins into
clathrin-coated vesicles
Delivery of Rab to a membrane
GDI: GDP-dissociation inhibitor
Rab is carried in the cytosol bound to
a GDI, keeping it in the Rab/GDP
state.
At a membrane, a non-specific GDI-
displacement factor can remove the
Rab/GDP from the GDI and insert it
into the membrane.
If a specific Rab-GEF is present, the
GDP on Rab will be exchanged for
GTP and the active Rab/GTP can
interact with effector proteins. If not,
the Rab/GDP will be removed by a
GDI and carried to another
membrane.
Vesicle fusion
Vesicle fusion is initiated by Rab/GTP, and
continues with SNARE-SNARE interaction.
Interaction provides energy to bring the vesicle
and target membranes into close proximity.
This proximity destabilizes the lipid bilayers,
and the vesicle and target membrane fuse.
Fusion recruit NSF and SNAP.
The NSF/SNAP complex disassemble the
SNARE complex using energy from the
hydrolysis of ATP.
GDI: GDP-dissociation inhibitor
Lysosomes
Electron micrograph of lysosomes and mitochondria in a mammalian cell
Lysosomes are membrane-enclosed organelles that contain an array of enzymes
capable of breaking down all types of biological polymers, proteins, nucleic acids,
carbohydrates, and lipids.
Yeasts and plant cells lack
lysosomes.
Vacuole assume the functions of
lysosomes in these cells as well
as performing other tasks.
In contrast to lysosomal
targeting, proteins are directed to
vacuoles by short peptide
sequences instead of by
carbohydrate markers.
Plant cell vacuole functions as lysosome in
addition to
Storing nutrients
Maintaining osmotic balance
Most lysosomal enzymes are acid
hydrolases, which are active at the acidic
pH.
The acidic internal pH results from the
action of a proton pump in the lysosomal
membrane, which imports protons from the
cytosol coupled to ATP hydrolysis.
Lysosomal storage diseases
Gaucher disease
Defficient in enzyme required
for the breakdown of glycolipids
I-cell disease
Defficient in enzyme that tags
lysosomal enzymes with mannose-6-
phosphate in Golgi apparatus
Exocytosis
Exocysts are complexes of 8 different proteins
formed during exocytosis.
Tethering and docking at exocysts results in
normal SNARE-mediated vesicle fusion.
Endocytosis
The acid hydrolases dissociate
from the mannose-6-phosphate
receptors when the transport
vesicles fuse with late
endosomes, and the mannose-6-
phosphate receptors are recycled
back to the trans Golgi network.
Phagocytosis and Autophagy
Autophagy is activated during
Nutrient starvation of the cell
It allows cells to degrade nonessential proteins and organelles so that
their components can be reutilized.
Developmental processes of multicellular organisms
For example: insect metamorphosis, which involves extensive tissue
remodeling and degradation of cellular components.
Programmed cell death
Defects in autophagy have been linked to several human diseases, in this
regards, including neurodegenerative diseases and cancer.