Podcast
Questions and Answers
What primarily determines the specific functions of different cellular membranes?
What primarily determines the specific functions of different cellular membranes?
- The overall size and shape of the organelles they enclose.
- The types of carbohydrates present on the membrane surface.
- The ratio of cholesterol to phospholipids.
- The specific asymmetry and composition of lipids. (correct)
How does the composition of lipids in the plasma membrane contribute to membrane asymmetry?
How does the composition of lipids in the plasma membrane contribute to membrane asymmetry?
- By symmetrically arranging cholesterol to stabilize both leaflets equally.
- By dynamically changing the lipid composition in response to external stimuli, ensuring symmetry over time.
- By ensuring an equal distribution of all lipid types between the inner and outer leaflets.
- By localizing specific lipids to either the inner or the outer leaflet, creating distinct domains. (correct)
Which of the following mechanisms is primarily responsible for maintaining the asymmetric distribution of lipids in cellular membranes?
Which of the following mechanisms is primarily responsible for maintaining the asymmetric distribution of lipids in cellular membranes?
- Uniform synthesis and insertion of lipids into both leaflets of the membrane.
- The high concentration of cholesterol that stabilizes the lipid bilayer equally on both sides.
- Spontaneous diffusion of lipids across the membrane.
- The action of flippases and floppases, which selectively move lipids across the bilayer. (correct)
Within a cell, how do organelles maintain their unique lipid compositions?
Within a cell, how do organelles maintain their unique lipid compositions?
What is the primary role of clathrin-coated vesicles in cellular trafficking?
What is the primary role of clathrin-coated vesicles in cellular trafficking?
What role do adaptins play in clathrin-mediated endocytosis?
What role do adaptins play in clathrin-mediated endocytosis?
Which protein is directly responsible for constricting and pinching off clathrin-coated vesicles from the plasma membrane?
Which protein is directly responsible for constricting and pinching off clathrin-coated vesicles from the plasma membrane?
What is the primary function of the low pH environment in endosomes and lysosomes?
What is the primary function of the low pH environment in endosomes and lysosomes?
How does the acidification of endosomes contribute to the regulation of receptor-mediated endocytosis?
How does the acidification of endosomes contribute to the regulation of receptor-mediated endocytosis?
What is the role of vacuolar ATPases (V-ATPases) in endosomal and lysosomal function?
What is the role of vacuolar ATPases (V-ATPases) in endosomal and lysosomal function?
Which of the following is a typical pH range found within lysosomes?
Which of the following is a typical pH range found within lysosomes?
What type of enzymes are predominantly found within lysosomes?
What type of enzymes are predominantly found within lysosomes?
In receptor-mediated endocytosis, what happens to LDL after it is internalized and reaches the lysosome?
In receptor-mediated endocytosis, what happens to LDL after it is internalized and reaches the lysosome?
What is the primary function of the transferrin receptor?
What is the primary function of the transferrin receptor?
During the endocytosis of iron via the transferrin receptor, what triggers the release of iron from transferrin in the endosome?
During the endocytosis of iron via the transferrin receptor, what triggers the release of iron from transferrin in the endosome?
What typically happens to the transferrin receptor after it delivers iron to the endosome?
What typically happens to the transferrin receptor after it delivers iron to the endosome?
How does endocytosis of growth factor receptors, such as the epidermal growth factor (EGF) receptor, contribute to the regulation of cell signaling?
How does endocytosis of growth factor receptors, such as the epidermal growth factor (EGF) receptor, contribute to the regulation of cell signaling?
What are the primary destinations for endocytosed receptors after they enter the endosomal system?
What are the primary destinations for endocytosed receptors after they enter the endosomal system?
What distinguishes phagocytosis from other forms of endocytosis?
What distinguishes phagocytosis from other forms of endocytosis?
Which cell types are primarily responsible for carrying out phagocytosis in mammals?
Which cell types are primarily responsible for carrying out phagocytosis in mammals?
What is the primary purpose of autophagy in cells?
What is the primary purpose of autophagy in cells?
Which cellular structure is primarily involved in the process of autophagy?
Which cellular structure is primarily involved in the process of autophagy?
Following the formation of an autophagosome, what is its next destination inside the cell?
Following the formation of an autophagosome, what is its next destination inside the cell?
Which of the following best describes the path of proteins from the endoplasmic reticulum (ER) to the extracellular space?
Which of the following best describes the path of proteins from the endoplasmic reticulum (ER) to the extracellular space?
What distinguishes constitutive secretion from regulated secretion?
What distinguishes constitutive secretion from regulated secretion?
Which of the following proteins is typically released via regulated secretion?
Which of the following proteins is typically released via regulated secretion?
How is insulin processed from its initial precursor form to its mature form?
How is insulin processed from its initial precursor form to its mature form?
What is the role of the C-peptide in insulin production?
What is the role of the C-peptide in insulin production?
Pepsinogen is secreted by chief cells and converted to its active form, pepsin, in the stomach lumen. What triggers the activation of pepsinogen?
Pepsinogen is secreted by chief cells and converted to its active form, pepsin, in the stomach lumen. What triggers the activation of pepsinogen?
What is the relationship between membrane traffic dysregulation and disease?
What is the relationship between membrane traffic dysregulation and disease?
Which protein is affected in cystic fibrosis, leading to defects in chloride ion transport?
Which protein is affected in cystic fibrosis, leading to defects in chloride ion transport?
How does the mutation in CFTR typically affect its function and localization?
How does the mutation in CFTR typically affect its function and localization?
What is the underlying cause of Niemann-Pick Type C (NP-C) disease?
What is the underlying cause of Niemann-Pick Type C (NP-C) disease?
To maintain cellular homeostasis, how do cells ensure that cholesterol is transported efficiently?
To maintain cellular homeostasis, how do cells ensure that cholesterol is transported efficiently?
Familial hypercholesterolemia is linked to defects in which of the following processes?
Familial hypercholesterolemia is linked to defects in which of the following processes?
What role do apolipoproteins play in lipid transport?
What role do apolipoproteins play in lipid transport?
How does the asymmetry of lipids in cellular membranes primarily impact cellular function?
How does the asymmetry of lipids in cellular membranes primarily impact cellular function?
What is the significance of the pH difference between the endosome and the cytosol?
What is the significance of the pH difference between the endosome and the cytosol?
What is the role of glycosidases within the lysosome?
What is the role of glycosidases within the lysosome?
How does the endocytosis of the Epidermal Growth Factor Receptor (EGFR) contribute to the regulation of cell growth and proliferation?
How does the endocytosis of the Epidermal Growth Factor Receptor (EGFR) contribute to the regulation of cell growth and proliferation?
How does the disruption of CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) trafficking lead to cystic fibrosis?
How does the disruption of CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) trafficking lead to cystic fibrosis?
Flashcards
Membrane Traffic
Membrane Traffic
The movement of substances into, within, and out of cells.
Endocytosis
Endocytosis
Uptake of extracellular nutrients, iron transport, protein degradation, and signal regulation are examples of this process
Endocytosis via Clathrin
Endocytosis via Clathrin
A clathrin-coated vesicle is part of this process
Dynamin
Dynamin
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Clathrin-mediated Endocytosis
Clathrin-mediated Endocytosis
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Pinocytosis
Pinocytosis
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Phagocytosis
Phagocytosis
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Lysosome
Lysosome
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Autophagy
Autophagy
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Receptor-Mediated Endocytosis
Receptor-Mediated Endocytosis
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Transferrin receptor
Transferrin receptor
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LDL
LDL
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LDL receptor
LDL receptor
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EGF Receptor
EGF Receptor
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Exocytosis
Exocytosis
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Lipoproteins
Lipoproteins
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Apoproteins
Apoproteins
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pH 4.5
pH 4.5
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Extracellular
Extracellular
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Cystic Fibrosis
Cystic Fibrosis
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CFTR
CFTR
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Niemann-Pick Type C
Niemann-Pick Type C
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Cytosol
Cytosol
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Cytoplasm
Cytoplasm
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Study Notes
- PH1123 is about membrane traffic
Membrane Lipids
- Study the varieties, organization, and membrane proteins associated with membrane lipids
Membrane Traffic
- Focus on endocytosis and secretion (exocytosis)
- Study the link to membrane traffic and disease
- Materials should be reviewed alongside other PH1123 content and Workshop material
- Essential Cell Biology by Alberts is a suggested reading material
Cell Structure Reminder
- Cells are surrounded by plasma membrane
- Cells contain several intracellular membrane-bound compartments
- It is important consider how many organelles in a typical mammalian cells are surrounded by a lipid bilayer.
Lipids
- Lipids are found in cellular membranes
- Types of lipids include:
- Cholesterol
- Glycolipids
- Phosphatidylcholine
- Sphingomyelin
- Phosphatidylethnolamine
- Phosphatidylinositol
- Phosphatidylserine
- Phosphatidylglycerol
- Phosphatidic acid
Membrane Proteins
- Proteins are on the membrane, not just the plasma membrane
- Types of membrane proteins include the following:
- Transmembrane
- Membrane-associated
- Lipid-linked
- Protein-attached
- Drug targets include receptors and channels
Organization of Lipids
- Lipids organize asymmetrically on membranes
- Phospholipid composition and distribution varies in plasma membranes
- Different organelles have different lipid compositions
Intracellular Transport
- Material moves into, within, and out of cells
Endocytic Pathways
- Types of endocytic pathways include the following:
- Pinocytosis
- Macropinocytosis
- Phagocytosis
- Clathrin-dependent endocytosis
- Caveolin-dependent endocytosis
- Clathrin and caveolin-independent pathways
Endocytosis
- Endocytosis is the internalization of molecules
- The cell uptakes extracellular nutrients like iron-transferrin via transferrin receptor
- The cell uptakes fats-LDL, via LDL receptor
- Proteins are endocytosed for degradation and down regulation of oncogenic signals, and therefore, for cancer
- Endocytosis involved cell recovery through endocytic recycling of proteins and lipids
- Endocytosis is involved in cholesterol homeostasis via LDL receptor
- Clathrin-coated vesicles mediate endocytosis
- Material is taken into cells specifically in clathrin-coated vesicles
- Coat proteins, clathrin, and adaptins play a role in endocytosis
Endocytosis and pH
- Extracellular pH is 7.4
- Early Endosome pH is 6.5 (2-5 min)
- Late endosome pH is 5.5-4.5 (10 min)
- Lysosome pH 5.5-4.5
- Vacuolar ATPase is a proton pump, which drops the pH
- Lysosomes and late endosomes act as the cell's "stomach"
- Lysosomes harbor enzymes, such as Nucleases, Proteinases, Lipases, and Glycosidases
- These enzymes have a pH of 4.5
Receptor-Mediated Endocytosis
- All receptor-mediated endocytosis relies on clathrin-mediated endocytosis
- Transferrin binds to transferrin receptor in this process
- Epidermal growth factor (EGF) binds to EGF Receptor in this process
- LDL binds to LDL Receptor in this process
- MPharm Degree- Receptors act as targets for drugs
Iron
- Cellular iron is needed
- Iron functions in cells by endocytosis
- Iron deficiency can cause anaemia (too low)
- Iron overload can cause Haemochromatosis (too high)
Iron Delivery
- Transferrin is an iron-currying protein
- Transferrin receptor
- Clathrin-coated vesicle uncoats
- Fusion occurs
- Recycling endosome
- Degradation or Downregulation of late endosomes or lysosomes
EGF Endocytosis
- Receptor-mediated endocytosis of epidermal growth factor (EGF) occurs by:
- EGF receptor and EGFR binding
- Clathrin-coated vesicle
- Signalling
- Fusion
- Endosomes
- Growth & Cell Division
- Cancer
- Degradation
- Downregulation of receptor and signalling
Cell Signaling
- The life of the cell is complex
Arwyn Jones Workshop
- This workshop will cover the following questions:
- What is the link between EGFR, HER2, HER3, and HER4?
- What is the link between HER2 and Breast Cancer?
- What is Herceptin aka Trastuzumab?
- What is an ADC, and what is Kadcyla?
Lipid Transportation
- With endogenous fats, fats that are hydrophobic need specialized transport through Lipoprotein complexes.
- Products of digestion are carried by proteins
- Triglycerides are carried via B-globulins
- Cholesterol* is carried via B-globulins and is synthesised in body
- Phospholipids are carried via a-globulins
- Free fatty acids are carried via albumins
- Plasma lipids and lipid transport are insoluble, but need to be transported across the membrane as follows:
- For solubility
- For recognition
- FFAs are bound to the plasma protein albumin
- Cholesterol, triglycerides, and phospholipids are transported as lipoprotein complexes
- Proteins associated with them are called APOPROTEINS
- Major apoproteins - APO E, APO C, APO B
- LDL- very important lipoprotein complex
Animations
- animations explain the link between Familial Hypercholesterolaemia and membrane traffic
- http://www.youtube.com/watch?v=Yanklj65zjs&feature=related
- http://www.youtube.com/watch?v=tNhESIfWxLc&feature=related
LDL Receptor
- Receptor-mediated endocytosis of LDL Receptor transports free cholesterol
- Includes the following:
- LDL receptor
- Clathrin-coated vesicle uncoating and fusion
- Recycling endosome
- Lysosome Degradation
Phagocytosis
- Phagocytosis involves cell eating from the outside
- Immune cells, such as macrophages, neutrophils, and dendritic cells use this process
- 1011 red blood cells/day are degraded by phagocytosis
Autophagy
- Autophagy means eating from the inside
- Old and damaged organelles are wrapped in membrane
- Then they are delivered to lysosomes for degradation
Additional Reading & Objectives
- You should be familiar with organelles that are surrounded by lipid membranes
- What is the difference between the cytoplasm and cytosol?
- Asymmetry of plasma membrane lipids
- Types of lipids found on membranes, and types of membrane proteins
- Compare endocytosis and Endocytic pathways
- Discuss the similarities and differences between endocytosis of the transferrin receptor, the EGF receptor and LDL receptor
- Focus on enzymes in the lysosomes, the role of iron in cells, downregulation of oncogenic signals, Apolipoproteins and disease, and LDL receptor biology
- Autophagy: learn what autophagy is
Secretory Pathways & Disease
- Membrane Traffic 1: Endocytosis involved the following
- Pinocytosis
- Phagocytosis
- With clathrin-dependent endocytosis
- Autophagy
The Cellular Soup
- II- how does material move out of cells
Exocytosis
- Exocytosis is the secretory pathway
Delivery of Membrane proteins
- Transferrin receptor
- CFTR- see later
- LDL Receptor
Exocytosis
- Exocytosis releases soluble proteins
- For example: Insulin (hormone), pepsinogen (proenzyme)
- Small molecules via exocytosis
- For example: Neurotransmitters
- acetylcholine
- noradrenaline
- Proteins travel from the ER to the extracellular fluid in vesicles
Transport
- Retrograde versus Anterograde
Insulin
- Insulin is synthesised using recombinant DNA technology in bacteria E.Coli
- Hormone is discovered to be processed as a larger immature form
- Hormone isolated in form that could be administered therapeutically to humans
- Hormone have its MOA elucidated
- Hormone determined for its primary and secondary structure
- Hormone to be measured using radioimmunoassay
- Intracellularly, preprotein Convertase aka signal peptidase gives signal sequence and preproinsulin in ER
- Carboxypeptidases gives C-peptide and mature insulin - Occurs in secretory granules
Insulin Production
- Insulin is packaged, budded, processed, and then goes through fission
- Ribosomes helps preprotein Convertases to give signal sequence and preproinsulin in ER
- Signal Peptidase then cleave signal peptide in ER
Pepsinogen
- Pepsinogen undergoes exocytosis and plays a role in Physiology
- Pepsinogen release from secretory granules is released by Gastrin Stimulation by:
- Food in Stomach and Vagal Stimulation
- Acid Secretion are discussed at the Workshop
- This slide on amended form better highlights the Chief cell is secreting pepsinogen
Pepsinogen secretion
- Pepsinogen is packaged, budded, and released during Fission at the TGN, then the ER by Ribosomes
- In Sectretory Granules, pH is 6
- Site is Active but INACTIVE
- there is Activation Peptide
- In Stomach Lumen, pH is les than 6.
- Therefore it is ACTIVE
Disease & Membrane Traffic
- Many 100s of diseases are associated with membrane traffic
- Cystic fibrosis - indirect
- The traffic of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR).
- A workshop is held with discussion around this
- Niemann Pick Type C (NP-C) - direct
- Defects occur in transport of lipids to lysosomes
- A Lysosomal Storage Disease
Cystic Fibrosis
- The CFTR is normally delivered by membrane traffic to the plasma membrane
- CFTR functions as a chloride channel
- In case of mutation, Transport is blocked and CFTR mutant accumulates. Excellent Video cover Cystic Fibrosis regarding function and cause: https://www.youtube.com/watch?v=_j99-xgOlaw
- The CFTR transports blocked at the Endoplasmic reticulum
Learning Objectives
- You should be familiar with:
- Organelles that are surrounded by lipid membranes
- What is the difference between the cytoplasm and cytosol?
- Asymmetry of plasma membrane lipids
- Types of lipids found on membranes- types of membrane proteins
- How cholesterol sits on the plasma membrane; why is it there?
- Endocytosis and Endocytic pathways
- Similarities and differences between endocytosis of the transferrin receptor, the EGF receptor and LDL receptor
- Enzymes in the lysosomes, role of iron, Apolipoproteins, LDL receptor biology, etc.
- Autophagy and the secretory pathway
- Focus on insulin Pepsinogen secretion and activation
- Topics include the following:
- membrane traffic
- cytoskeleton
- acid secretion
- Antibody Drug Conjugates?
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