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What are the two main types of lipid constituents in biomembranes?
What are the two main types of lipid constituents in biomembranes?
Phospholipids and sphingolipids.
What is the main function of membranes?
What is the main function of membranes?
Phospholipids have a _____ head group and _____ tails.
Phospholipids have a _____ head group and _____ tails.
polar, non-polar
All lipids found in human plasma have similar structures.
All lipids found in human plasma have similar structures.
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What does the fluid-mosaic model describe?
What does the fluid-mosaic model describe?
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What is the role of receptors in membranes?
What is the role of receptors in membranes?
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Which technique confirmed the bilayer structure of membranes?
Which technique confirmed the bilayer structure of membranes?
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What are spherocytes?
What are spherocytes?
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What do we mean by integral and peripheral?
What do we mean by integral and peripheral?
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Proteins in the membrane are completely immobilized.
Proteins in the membrane are completely immobilized.
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How do red blood cells (RBCs) maintain their shape?
How do red blood cells (RBCs) maintain their shape?
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What role does spectrin play in erythrocytes?
What role does spectrin play in erythrocytes?
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Which of the following is true about lipid rafts?
Which of the following is true about lipid rafts?
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What is one characteristic of cholesterols in membranes?
What is one characteristic of cholesterols in membranes?
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What does FRAP stand for?
What does FRAP stand for?
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What are the two types of proteins based on their membrane association?
What are the two types of proteins based on their membrane association?
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Cholesterol is essential to the process of raft formation.
Cholesterol is essential to the process of raft formation.
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Match the following terms with their definitions:
Match the following terms with their definitions:
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What are GPI-linked proteins?
What are GPI-linked proteins?
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The process of membrane proteins moving laterally within the bilayer is referred to as ______.
The process of membrane proteins moving laterally within the bilayer is referred to as ______.
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What is required for transverse diffusion of lipids?
What is required for transverse diffusion of lipids?
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What is the role of annexin V in apoptosis?
What is the role of annexin V in apoptosis?
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What are the two hypotheses for maintaining membrane asymmetry?
What are the two hypotheses for maintaining membrane asymmetry?
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What does an activation of scramblase cause?
What does an activation of scramblase cause?
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Phosphatidylserine (PS) at the outer leaflet indicates a healthy cell.
Phosphatidylserine (PS) at the outer leaflet indicates a healthy cell.
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The process by which phospholipids flip from one leaflet to another in a membrane is called ______.
The process by which phospholipids flip from one leaflet to another in a membrane is called ______.
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Which molecules are translocated rapidly to the inner leaflet in ATP-replete erythrocytes?
Which molecules are translocated rapidly to the inner leaflet in ATP-replete erythrocytes?
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What is the conclusion regarding the transport of PS and PE?
What is the conclusion regarding the transport of PS and PE?
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What is the role of phospholipase A2 in the outer leaflet?
What is the role of phospholipase A2 in the outer leaflet?
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How can the activity of ATP-dependent amino-phospholipid translocase be measured?
How can the activity of ATP-dependent amino-phospholipid translocase be measured?
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What does FRAP stand for?
What does FRAP stand for?
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A student should use fluorescently-tagged analogues that are more polar than the target phospholipid for better incorporation.
A student should use fluorescently-tagged analogues that are more polar than the target phospholipid for better incorporation.
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Why is it important to study membrane proteins?
Why is it important to study membrane proteins?
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What are the two main types of membrane proteins?
What are the two main types of membrane proteins?
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Integral membrane proteins are easier to remove than peripheral proteins.
Integral membrane proteins are easier to remove than peripheral proteins.
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Detergents disrupt the _____ bilayer to solubilize membrane proteins.
Detergents disrupt the _____ bilayer to solubilize membrane proteins.
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What forces hold peripheral proteins to the membrane?
What forces hold peripheral proteins to the membrane?
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What is the role of Triton X-100 in the protocol described?
What is the role of Triton X-100 in the protocol described?
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What is the purpose of performing hydropathy plots?
What is the purpose of performing hydropathy plots?
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Why are membrane proteins challenging to study?
Why are membrane proteins challenging to study?
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What class of signaling allows for long-distance communication between cells?
What class of signaling allows for long-distance communication between cells?
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A good signal must be able to travel from site of manufacture to the _____ site.
A good signal must be able to travel from site of manufacture to the _____ site.
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What does Kd approximate?
What does Kd approximate?
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What is the Kd for the hepatocyte insulin receptor for insulin?
What is the Kd for the hepatocyte insulin receptor for insulin?
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What is the normal insulin concentration in blood?
What is the normal insulin concentration in blood?
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What percentage of receptors will be bound to insulin at a normal concentration of 5 x 10^-12 M?
What percentage of receptors will be bound to insulin at a normal concentration of 5 x 10^-12 M?
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If the normal circulating insulin concentration was 1.4 x 10^-10 M, what percentage of receptors are bound to insulin?
If the normal circulating insulin concentration was 1.4 x 10^-10 M, what percentage of receptors are bound to insulin?
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What happens if insulin concentration is increased 5-fold from 1.4 x 10^-10 M?
What happens if insulin concentration is increased 5-fold from 1.4 x 10^-10 M?
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What is the formula for calculating Fractional Occupancy (F.O.)?
What is the formula for calculating Fractional Occupancy (F.O.)?
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What neurotransmitter is released from pre-ganglionic fibers?
What neurotransmitter is released from pre-ganglionic fibers?
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What are the principal secretory products of chromaffin cells?
What are the principal secretory products of chromaffin cells?
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How long is the half-life of circulating adrenaline?
How long is the half-life of circulating adrenaline?
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Adrenaline has a longer lasting effect in the body compared to its half-life.
Adrenaline has a longer lasting effect in the body compared to its half-life.
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What causes adrenaline to have different effects in different tissues?
What causes adrenaline to have different effects in different tissues?
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What are the three crucial criteria for a good receptor?
What are the three crucial criteria for a good receptor?
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How does the Kd value relate to receptor-ligand binding?
How does the Kd value relate to receptor-ligand binding?
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The binding affinity of adrenaline is high due to its low Kd.
The binding affinity of adrenaline is high due to its low Kd.
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What experimental method is used to detect weak binding of a ligand to its receptor?
What experimental method is used to detect weak binding of a ligand to its receptor?
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Study Notes
Introduction to Biomembranes
- Biomembranes consist of different classes of lipids, influencing their physical properties.
- Basic structure was determined through various experiments, including freeze-etching and electron microscopy.
- Original fluid-mosaic model has evolved with new findings about membrane dynamics.
Functions of Membranes
- Compartmentation: Enables separation of enzymes, metabolites, and pathways within cells.
- Specific Transport: Regulates passage of compounds, either passively or actively (e.g., pumps).
- Maintenance of Electrical Potential: Membranes can create and sustain electrical gradients.
- Energy Trapping: Some membranes convert light energy into electrical or chemical energy.
- Signal Recognition/Transduction: Membranes have receptors that interact with hormones and neurotransmitters, generating second messengers.
- Shape Maintenance: The cytoskeleton helps maintain cell shape and structure through membrane interaction.
- Endocytosis/Exocytosis: Membrane trafficking is a constant process facilitating material exchange.
Phospholipids as Main Lipids
- Phospholipids include hydrophilic head groups and hydrophobic fatty acyl tails, resulting in amphipathic properties.
- Classes of lipids in biomembranes:
- Phosphoglycerides: Examples include PC, PE, PS, and PI.
- Sphingolipids: Derivatives like sphingomyelin with complex structures.
- Sterols: Cholesterol, an important component affecting membrane fluidity.
Lipid Composition and Diversity
- Over 500 lipid species have been identified in human plasma, showcasing diversity.
- Fatty acids can vary in chain length and saturation (saturated vs. unsaturated).
- Micelles are rare; natural phosphoglycerides are too bulky for micelle formation.
Membrane Structure Discovery
- Robertson's hypothesis in the 1950s linked the appearance of membranes to osmium tetroxide binding.
- Techniques like freeze-etching revealed bilayer structures and protein distributions within membranes.
Membrane Proteins and Mobility
- Protein Distributions: Inner and outer leaflets display different protein densities; proteins play crucial roles in signaling and transport.
- Phospholipids and proteins are mobile within the bilayer, allowing lateral diffusion (within the same plane); transverse diffusion (flipping) is less common and ATP-dependent.
Experimental Evidence of Lateral Diffusion
- Human-mouse heterokaryon experiments showed mobility of proteins was unaffected by ATP depletion or temperature changes, demonstrating lateral diffusion.
Red Blood Cell Shape and Structure
- Spherocytes are abnormally shaped red blood cells resulting from defects in cytoskeletal proteins (e.g., spectrin, ankyrin).
- RBC shape is maintained by a network of integral and peripheral membrane proteins associated with the cytoplasmic face.
Types of Membrane Proteins
- Integral Proteins: Span across both layers of the membrane, involved in transport and signaling functions.
- Peripheral Proteins: Associated with one side of the membrane; often involved in structural support and signaling.### Membrane Protein Mobility in Mice
- Comparison of lateral mobility of membrane proteins in normal mice and spherocytic mice (lacking spectrin).
- Sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE) employed on erythrocyte “ghosts” to analyze protein mobility.
- Spherocytic red blood cells (RBCs) confirmed via RB2 labeling.
- Fluorescent labeling techniques used: DTAF (Dichlorotriazinyl aminofluorescein) and Concanavalin A (Con A).
- Fluorescence recovery after photobleaching (FRAP) utilized to evaluate diffusion rates of labeled proteins in RBCs.
Lateral vs Transverse Mobility
- Lateral diffusion in erythrocytes is rapid compared to transverse diffusion.
- Transverse diffusion for lipids requires significant time (1-2 hours), while lateral movement is much faster.
- Proteins embedded in membranes typically do not exhibit transverse "flipping."
Micro-Domains and Bilayer Asymmetry
- Membrane micro-domains are noted, highlighting non-random distributions of proteins.
- Spectrin "corrals" influence erythrocyte membrane organization.
- Lipid asymmetry established between inner and outer leaflets of the bilayer.
- GPI (Glycophosphatidylinositol)-anchored proteins exhibit unique trafficking and stability characteristics.
GPI-Linked Proteins
- GPI-linked proteins demonstrate poor solubility in non-ionic detergents.
- Breakdown of protein-lipid interactions occurs under harsher conditions.
- High fractions of sphingolipids found in insoluble fractions of membranes.
Lipid Raft Model
- Lipid rafts create "liquid-ordered" regions influenced by sphingolipid and cholesterol composition.
- Rafts facilitate the retention and trafficking of specific proteins.
- Stability of rafts enhanced through hydrogen bonding with hydroxyl groups in sphingolipids.
Discovery Techniques
- Atomic force microscopy reveals distinct ordering of lipid rafts.
- Rafts contribute to varied membrane composition between cell types.
Membrane Dynamics
- Fluid-mosaic model updated to incorporate stability provided by cytoskeletal interactions.
- Non-random distribution of membrane proteins leads to functional implications.
Transport Mechanisms
- Active transport mechanisms, including flippases, floppases, and scramblases, maintain lipid asymmetry.
- Apoptosis triggers changes in PL distribution, with phosphatidylserine (PS) exposure indicating cell death.
Important Concepts in Apoptosis
- Apoptosis leads to morphological changes and altered lipid asymmetry, including the externalization of PS.
- Rapid translocation of *PS and *PE to the inner leaflet under ATP-rich conditions exemplifies facilitated transport mechanisms essential for maintaining membrane integrity.### Cell Differentiation and Apoptosis
- Differentiation of fingers and toes in a human embryo occurs due to apoptosis of cells between digits.
- Phosphatidylserine (PS) is an important indicator of apoptosis.
Membrane Proteins Overview
- Integral and peripheral proteins are distinct types of membrane proteins, with integral proteins spanning the lipid bilayer.
- Protein asymmetry is absolute, meaning different proteins exist on inner and outer leaflets of membranes.
Mechanisms of Membrane Asymmetry
- Maintaining lipid asymmetry requires ATP.
- Two hypotheses exist regarding phospholipid translocation: slow "flip-flop" rate versus specific mechanisms maintaining inner leaflet phospholipids.
- Assessing asymmetry can be done through experiments measuring "flip-flop" rates using fluorescent or radiolabeled phospholipids.
Experimental Techniques
- Methods include using phospholipid vesicles and incubating erythrocytes with fluorescent phospholipid analogues to measure translocation rates.
- FRAP (Fluorescence Recovery After Photobleaching) can assess lateral diffusion versus transverse flipping.
Phosphatidylserine and Experimental Design
- For ATP-dependent amino-phospholipid translocase activity, choose fluorescently-tagged analogues that closely mimic unmodified phosphatidylserine.
- Understanding the polarity of analogues can influence incorporation into membranes and affect experimental outcomes.
Importance of Membrane Proteins
- Membrane proteins constitute over 30% of cellular proteins and are common targets for FDA-approved drugs, despite challenges in structural determination.
- RBCs are frequently studied for insights into membrane protein functionality due to their accessible membrane structure.
Anatomy of Membrane Proteins
- Integral membrane proteins feature transmembrane domains, disulfide bridges, and oligosaccharides attached in the Golgi and ER.
- The solubilization process for membrane proteins involves detergents that disrupt the lipid bilayer, forming protein-lipid-detergent complexes.
Reconstituting Membrane Proteins
- Reconstituting peripheral proteins requires different methods than those used for integral proteins, emphasizing context-specific techniques to isolate target proteins.
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Description
This quiz covers the introduction to biomembranes, focusing on their composition, structure, and physical properties. It highlights the development of the fluid-mosaic model and includes key experiments that shaped our understanding of biomembranes. Ideal for students exploring cell signalling and membrane science.