Lecture 14 Vesicular Transport II Flashcards
what are the membrane compartments of lysosome filled with
hydrolytic enzymes
what is the function of lysosomes
intracellular digestion of macromolecules
what type of enzymes are found in lysosomes
proteases nucleases gylcosidases lipases phospholipase phosphates sullfatases
what do lysosomes require for optimal activation
acidic environment
proteolytic cleavage
vacuolar ATPase
pumps H+ into lysosomes to maintain acidic pH and drive transport of small metabolites
what does the Trans-Golgi network deliver to lysosomes
membrane proteins and hydrolyses
steps in co-translation transport of membrane proteins and hyrolases to lysosome in TGN
into rough Er
then transported via Golgi comes to TGN
TGN buds off Golgi to form
endosomes
these develop into lysosomes
what do lysosomal hydrolyses have that attach them in the CGN
sorting signal
sorting signal of lysosomal hydrolyses
Mannose 6 Phosphate
what do M6P receptors in TGN recognize
sorting signal sugar, M6P
after hydrolyses bind to TGN…
they are packaged into clathrin coated vesicles that bud off and delivered to endosomes then lysosomes
steps of lysosomal protein sorting
transport of newly synthesized lysosomal hydrolases to lysosomes
- on cis-Golgi mannose binds to hydrolase from ER and phosphate is added – becomes M6P
- M6P signal is uncovered as it moves thru Golgi
- M6P binds to M6P receptor, gets clathrin coated and buds off
- Transport to endosome, fusion with endosome
- hydrolyses dissociate from M6P receptors (due to low pH)
- Empty transports are recycled back to the TGN
- in endosomes, phosphate is removed and hydrolyses are kept in the endosome
what is the enzyme responsible for recognizing the lysosomal hydrolase?
GlCNAc phosphotransferase
how does the lysosome recognize the hydrolase
signal patch binds to the phsphotransferase
binding site of phosphotransferase binds
high mannose N-linked oligosaccharides
UDP-GLCNAc
steps in lysosomal recognition
- binding of signal path to recognition site of phosphotransferase
- transfer of GlcNAc-P to mannose at catalytic site
- release
- remold of GLcNAc
lysosomal storage disease
caused by genetic defects in lysosomal hydrolyses
results in accumulation of undigested material in lysosome
Inclusion Cell disease
Hurlers Disease
Missing GlcNAc phosphtransferase
Hurlers disease
mutation in the enzyme required to break down glycosaminoglycan chains
lysosomal storage disease
Inclusion Cell Disease
missing hydrolyses in cell types
substrates accumulate as inclusions
missing GlcNAc phosphotransferases cause
enzymes that aren’t phosphorylated and therefore not sorted into vesicles and delivered into lysosomes but instead are carried to cell surface and found in blood
transport mechanisms in this lecture
- Transport from TGN to Lysosomes
- endocytosis
- exocytosis
endocytosis
uptake of macromolecules from exterior across plasma membrane
pinocytosis
form of endocytosis
small particles are ingested by pinocytic vesicles
phagocytosis
form of endocytosis
large particles are ingested by phagosomes
receptor mediated endocytosis
used to import macromolecules from outside cells
molecules bind to receptors on membrane surface then accumulate in clathrin coated pits
provides selective concentration mechanisms
what is an example of receptor mediated endocytosis
cholesterol uptake
blockage of this pathway results in atherosclerosis
role of clathrin coated pits in receptor mediated endocytosis
LDL receptors associate with clathrin-coated pits that then pinch off, carrying LDL particles and receptors
LDL particles are freed into extracellular space when clathrin is removed
steps of cholesterol uptake
- endocytosis
- uncaring
- fusion with endosome
- budding off of transport vesicles
- return of LDL receptors to plasma membrane
sequestering of endocytose proteins
- covalent modification with ubiquitin– requires only 1 ubiquitin
- ubiquitin tag facilitates the uptake of the receptors into endocytic vesicles
- ubiquitin is recognized by proteins that mediate the sorting process into the internal membrane vesicles of multi vesicular bodies
Lipid kinase helps regulate by phosphorylating phosphatidylinositol
serve as docking sites for the proteins that mediate the invagination proces
possible fates of endocytose receptor proteins
- recycled transport vesicles
- degradation by endolysosome
- transcytosis of transport vesicle
what type of cells use phagocytosis?
macrophages and neutrophils
phagosomes
macrophages and neutrophils use phagocytosis by
ingesting invading microorganisms to protect against infection
scavenge senescent and apoptotic cells
mechanism of phagocytosis
ex. antibody interaction
- antibodies bind to microbe
- Fc chain recognized by Fc receptor on surface of WBC
- pseudopod is formed and engulfs particle to form phagosome
what triggers phagocytosis?
binding of a particle to receptors on phagocyte surface
formation of pseudopod is driven by:
localized actin polymerization and reorganization
formation of pseudopod is controlled by
Rho GTPases
phosphoinostitide signaling
pinocytosis
cells ingest bits of PM in form of small pinocytotic vesicles
begins at clathrin coated vesicle
extraceullar fluid is trapped with solutes as they invaginate
flask shaped invaginations in PM
caveolae
caveolae
enriched in cholesterol and glycosphingolipids
caveoloae ingavingate into membrane by lipid composition
don’t connect with lysosomes
structural protein of caveolae
caveolin
exocytosis
transport from TGN to cell exterior
fusion of vesicles with PM
____ secretory pathway operates continuously
constitutive
____ secretory pathway is triggered by signals
regulated
signal combines with secretory vesicle and it is fused with membrane
three mechanisms of protein sorting in TGN
- signal mediated diversion to lysosomes
- signal mediated diversion to secretory vesicles
- constitutive secretory pathway
formation of secretory vesicles
involves selective aggregation/clumping of proteins
TGN promotes proteolytic processing of the proteins
steps in formation of synaptic vesicle
1, delivery of synaptic vesicle membrane components to presynaptic plasma membrane
2. endocytosis of synaptic vesicle membrane components to form new synaptic vesicles directly
3. endocytosis of synaptic vesicle membrane components \
and delivery to endosome
4. budding of synaptic from endosome
5. loading o fNeurotransmitter into synaptic vesicle
6. secret of Neurotransmitter by exocytosis