Membrane Trafficking
Membrane Trafficking
Membrane Trafficking
Compartmentalization of Cells
Membranes
Partition cell Important cellular functions Impermeable to most hydrophobic molecules contain transport proteins to import and export specific molecules Mechanism for importing and incorporating organelle specific proteins that define major organelles
Compartmentalization of Cells
Major Organelles
Nucleus
Cytosol ER Golgi Apparatus Mitochondria and Chloroplast Lysosomes Endosomes Peroxisomes
Cytosol Mitochondria Rough ER cisternae Smooth ER cisternae plus Golgi cisternae Nucleus Peroxisomes Lysosomes Endosomes
54 22 9 6 6 1 1 1
Organelles arising from pinching off of PM have interior equivalent to exterior of cell
Development of Mitochondria
Endosymbiosis of mitochondria and plastids creates double membrane organelle which have their own genome.
Development of plastids
Compartmentalization of Cells
3 Types of Transport Mechanisms
1. Gated Transport:
gated channels topologically equivalent spaces 2. Transmembrane Transport: protein translocators topologically distinct space 3. Vesicular transport:
Membrane Trafficking
Membrane Trafficking
Endo-membrane system General concepts of vesicle-mediated traffic Sequence of events beginning with ER and ending at PM Details of Golgi function Golgi to PM pathway The Endocytotic Pathway The Exocytotic Pathway
Endomembrane Network
The endomembrane system is a network of organelles in eukaryotic cells Exocytosis begins in the endoplasmic reticulum The Golgi apparatus modifies and sorts proteins in the exocytic pathway Exocytosis ends at the plasma membrane Endocytosis begins at the plasma membrane Endocytosis ends at the lysosome
extracellular
Ca2
Y Y
O2
Ca2
cytoplasm
Membrane lipids and proteins move to and from the plasma membrane during these processes
Vesicular Transport
RER to cis Golgi Modified in Golgi (glycosylation, phosphorylation) Sorted at trans Golgi network into
Lysosomal (endocytosis)
Regulated (exocytosis) constitutive (exocytosis)
In vesicle-mediated transport, a membrane-bound vesicle buds from one compartment and fuses with another.
The Golgi apparatus plays a central role in vesicular traffic within cells.
The Golgi apparatus modifies and sorts proteins in the exocytic pathway
The Golgi apparatus is organized into discrete compartments called cisternae. The cisternae are stacked on top of one another, and are classified as cis, medial, or trans according to their relative location within the overall Golgi structure. Golgi-resident proteins are primarily responsible for modifying proteins undergoing exocytosis. They are retained in the Golgi apparatus by transmembrane Golgi retention sequences.
The Golgi apparatus modifies and sorts proteins in the exocytic pathway
The extreme ends of the Golgi apparatus are elaborated into long, tubular structures called the cis Golgi network and trans Golgi network. Both Golgi networks sort proteins into vesicles targeted to different locations. The trans Golgi network is especially effective at sorting a large number of proteins into many distinct vesicle types.
Proteins exiting the ER join the Golgi apparatus at the cis Golgi network. The Golgi apparatus consists of a collection of stacked compartments.
Modification of the N-linked oligosaccharides is done by enzymes in the lumen of various Golgi compartments.
1. Sorting in TGN 2. Protection from protease digestion 3. Cell to cell adhesion via selectins
Once proteins that dont normally reside in the ER are properly folded, they are transported to the golgi apparatus.
Model for the formation of a clathrin-coated pit and the selective incorporation of integral membrane proteins into clathrin-coated vesicles
COPII coated vesicles transport via the vesicular tubular cluster (vtc) to the cis-Golgi network.
The protein coating is removed and the vesicles fuse with each other to form the vtc. The vtc is motored by kinesin (motor protein) along microtubules that function like engine on rail tracks. The vtc fuses with the cis-Golgi network. COPI transports vesicles back to ER
Large moving compartments that mature into the TGN, and return enzymes to trailing compartments by retrieval vesicles.
The trans Golgi network (TGN) sorts proteins exiting the Golgi apparatus
Exocytosis: a process of release of intracellular molecules (such as hormones, secretory proteins) contained within a membranebounded vesicle by fusion of the vesicle with its plasma membrane.
Endocytosis
Small region of the plasma membrane invaginates to form membrane-limited vesicles. Internalized molecules retain topology (lumen = extracellular) Cargo can be specifically selected (receptor) Destination of cargo can be controlled;
Pinocytosis
Extracellular fluid is captured in a vesicle and brought into the cell;
Receptor-mediated endocytosis
Specific molecules bind to surface receptors, which are then enclosed in an endocytic vesicle
Phagocytosis in Macrophage: The cells receptors in the plasma membrane enable them to recognize their targets. For example, macrophages have receptor that recognizes phosphatidylserine which becomes exposed on the surface of dead cells.
Classified as macropinocytosis (vesicles > 1 mm in diameter) and micropinocytosis (vesicles < 200 nm in diameter)
Receptor-Mediated Endocytosis
Nutrient Uptake (LDL,transferrin, etc.) Membrane Recycling Membrane Protein Recycling Antigen Uptake Synaptic Vesicle Recycling Signaling Receptor Down-Regulation
Receptors Bind Cargo. Clathrin Adaptins [AP1(Golgi) or AP2(PM)] bind to Receptor Tail Sequences. Coated Pits Form and Pinch Off into Coated Vesicles
Undergo endocytosis in response to a signal (ex. SV40 binding) in a cholesterol- and dynamin-dependent fashion Internalized caveolae recruit actin to form comet tails Upon internalization caveolae are delivered to novel endosomal compartments known as caveosomes
The endocytic pathway is divided into the early endosomes and late endosomes pathway.
Materials in the early endosomes are sorted:
Integral membrane proteins are shipped back to the membrane; Other dissolved materials and bound ligands retained. Dissociation of internalized ligand-receptor complexs in the late endosomes: Molecules that reach the late endosomes are moved to lysosomes.
One ultimate destination of some proteins that arrive in the TGN is the lysosome. These proteins include acid hydrolases.
Lysosomes are like the stomach of the cell. They are organelles surrounded by a single membrane and filled with enzymes called acid hydrolases that digest (degrade) a variety of macromolecules. A vacuolar H+ ATPase pumps protons into the lysosome causing the pH to be ~5.
The macromolecules that are degraded in the lysosome arrive by endocytosis phagocytosis or autophagy.
lysosomes
Autophagy
Exocytosis
Vesicle moves to cell surface Membrane of vesicle fuses Materials expelled orCell discharges material Reverse of endocytosis
Where do newly synthesized membrane and secretory proteins need to go and how do they get there?
Secretion (constitutive and regulated) PM protein delivery (polarized and non-polarized cells) Lysosomal targeting
Regulated secretion
Occurs in endocrine, exocrine and neuronal cells
Insulin secretion in pancreatic b-cells Trypsinogen secretion in pancreatic acinar cells
Figure 09.11: Transmission electron micrograph of clathrin-coated pits and vesicles at the oocyte surface.
Acidic pH of the gut favor association of antibody with Fc receptor whereas the neutral pH of the extracellular fluid favors dissociation.
Transcytosis: transport of macromolecular cargo from one side of the cell to the other Transcytosis is also utilized in the biosynthetic trafficking of some PM proteins pIgA-receptor is a model for studying transcytosis
Contains sorting information in its cytoplasmic tail pIgA is secreted into the the gut lumen, bile and milk as part of the mucosal immune response
pIgA-R
pIgA
Blood/interstitial