Biochem1 2014 Recitation Chapter 11 – Lipids/Membrane Structure
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Lipid Bilayer, with the Nonpolar Regions of the Lipids Facing Inward
Chapter 7 Membranes: Their Structure, Function, and Chemistry Lectures by Kathleen Fitzpatrick Simon Fraser University © 2012 Pearson Education, Inc. Membranes: Their Structure, Function, and Chemistry • Membranes define the boundaries of a cell, and its internal compartments • Membranes play multiple roles in the life of a cell © 2012 Pearson Education, Inc. Figure 7-1A © 2012 Pearson Education, Inc. Figure 7-1B © 2012 Pearson Education, Inc. The Functions of Membranes • 1. Define boundaries of a cell and organelles and act as permeability barriers • 2. Serve as sites for biological functions such as electron transport • 3. Possess transport proteins that regulate the movement of substances into and out of cells and organelles © 2012 Pearson Education, Inc. The Functions of Membranes (continued) • 4. Contain protein molecules that act as receptors to detect external signals • 5. Provide mechanisms for cell-to-cell contact, adhesion, and communication © 2012 Pearson Education, Inc. Figure 7-2 © 2012 Pearson Education, Inc. Models of Membrane Structure: An Experimental Approach • The development of electron microscopy in the 1950s was important for understanding membrane structure • The fluid mosaic model is thought to be descriptive of all biological membranes • The model envisions a membrane as two fluid layers of lipids with proteins within and on the layers © 2012 Pearson Education, Inc. Overton and Langmuir: Lipids Are Important Components of Membranes • In the 1890s Overton observed the easy penetration of lipid-soluble substances into cells and concluded that the cell surface had some kind of lipid “coat” on it • Langmuir studied phospholipids and found that they were amphipathic and reasoned that they must orient on water with the hydrophobic tails away from the water © 2012 Pearson Education, Inc. -
Biological Membranes and Transport Membranes Define the External
Biological Membranes and Transport Membranes define the external boundaries of cells and regulate the molecular traffic across that boundary; in eukaryotic cells, they divide the internal space into discrete compartments to segregate processes and components. Membranes are flexible, self-sealing, and selectively permeable to polar solutes. Their flexibility permits the shape changes that accompany cell growth and movement (such as amoeboid movement). With their ability to break and reseal, two membranes can fuse, as in exocytosis, or a single membrane-enclosed compartment can undergo fission to yield two sealed compartments, as in endocytosis or cell division, without creating gross leaks through cellular surfaces. Because membranes are selectively permeable, they retain certain compounds and ions within cells and within specific cellular compartments, while excluding others. Membranes are not merely passive barriers. Membranes consist of just two layers of molecules and are therefore very thin; they are essentially two-dimensional. Because intermolecular collisions are far more probable in this two-dimensional space than in three-dimensional space, the efficiency of enzyme-catalyzed processes organized within membranes is vastly increased. The Molecular Constituents of Membranes Molecular components of membranes include proteins and polar lipids, which account for almost all the mass of biological membranes, and carbohydrate present as part of glycoproteins and glycolipids. Each type of membrane has characteristic lipids and proteins. The relative proportions of protein and lipid vary with the type of membrane, reflecting the diversity of biological roles (as shown in table 12-1, see below). For example, plasma membranes of bacteria and the membranes of mitochondria and chloroplasts, in which many enzyme-catalyzed processes take place, contain more protein than lipid. -
Membrane Transport, Absorption and Distribution of Drugs
Chapter 2 1 Pharmacokinetics: Membrane Transport, Absorption and Distribution of Drugs Pharmacokinetics is the quantitative study of drug movement in, through and out of the body. The overall scheme of pharmacokinetic processes is depicted in Fig. 2.1. The intensity of response is related to concentration of the drug at the site of action, which in turn is dependent on its pharmacokinetic properties. Pharmacokinetic considerations, therefore, determine the route(s) of administration, dose, and latency of onset, time of peak action, duration of action and frequency of administration of a drug. Fig. 2.1: Schematic depiction of pharmacokinetic processes All pharmacokinetic processes involve transport of the drug across biological membranes. Biological membrane This is a bilayer (about 100 Å thick) of phospholipid and cholesterol molecules, the polar groups (glyceryl phosphate attached to ethanolamine/choline or hydroxyl group of cholesterol) of these are oriented at the two surfaces and the nonpolar hydrocarbon chains are embedded in the matrix to form a continuous sheet. This imparts high electrical resistance and relative impermeability to the membrane. Extrinsic and intrinsic protein molecules are adsorbed on the lipid bilayer (Fig. 2.2). Glyco- proteins or glycolipids are formed on the surface by attachment to polymeric sugars, 2 aminosugars or sialic acids. The specific lipid and protein composition of different membranes differs according to the cell or the organelle type. The proteins are able to freely float through the membrane: associate and organize or vice versa. Some of the intrinsic ones, which extend through the full thickness of the membrane, surround fine aqueous pores. CHAPTER2 Fig. -
Chapter 2 Cell Membranes
Chapter 2 Cell Membranes © 2020 Elsevier Inc. All rights reserved. Figure 2–1 The hydrophobic effect drives rearrangement of lipids, including the formation of bilayers. The driving force of the hydrophobic effect is the tendency of water molecules to maximize their hydrogen bonding between the oxygen and hydrogen atoms. Phospholipids placed in water would potentially disrupt the hydrogen bonding of water clusters. This causes the phospholipids to bury their nonpolar tails by forming micelles, bilayers, or monolayers. Which of the lipid structures is preferred depends on the lipids and the environment. The shape of the molecules (size of the head group and characteristics of the side chains) can determine lipid structure. (A) Molecules that have an overall inverted conical shape, such as detergent molecules, form structures with a positive curvature, such as micelles. (B) Cylindrical-shaped lipid molecules such as some phospholipids preferentially form bilayer structures. (C) Biological membranes combine a large variety of lipid molecular species. The combination of these structures determines the overall shape of the bilayer, and a change in composition or distribution will lead to a change in shape of the bilayer. Similarly a change in shape needs to be accommodated by a change in composition and organization of the lipid core. © 2020 Elsevier Inc. All rights reserved. 2 Figure 2–2 The principle of the fluid mosaic model of biological membranes as proposed by Singer and Nicolson. In this model, globular integral membrane proteins are freely mobile within a sea of phospholipids and cholesterol. © 2020 Elsevier Inc. All rights reserved. 3 Figure 2–3 Structure of phospholipids. -
Biological Membranes Transport
9/15/2014 Advanced Cell Biology Biological Membranes Transport 1 1 9/15/2014 3 4 2 9/15/2014 Transport through cell membranes • The phospholipid bilayer is a good barrier around cells, especially to water soluble molecules. However, for the cell to survive some materials need to be able to enter and leave the cell. • There are 4 basic mechanisms: 1. DIFFUSION and FACILITATED DIFFUSION 2. OSMOSIS 3. ACTIVE TRANSPORT 4. BULK TRANSPORT AS Biology, Cell membranes and 5 Transport 11.3 Solute Transport across Membranes 6 3 9/15/2014 Passive Transport Is Facilitated by Membrane Proteins Energy changes accompanying passage of a hydrophilic solute through the lipid bilayer of a biological membrane 7 Figure 11.2 Overview of membrane transport proteins. 4 9/15/2014 Figure 11.3 Multiple membrane transport proteins function together in the plasma membrane of metazoan cells. 5 9/15/2014 • Facilitated transport – Passive transport – Glucose – GLUT Cellular uptake of glucose mediated by GLUT proteins exhibits simple enzyme kinetics 11 12 6 9/15/2014 Regulation by insulin of glucose transport by GLUT4 into a myocyte 13 Effects of Osmosis on Water Balance • Osmosis is the diffusion of water across a selectively permeable membrane • The direction of osmosis is determined only by a difference in total solute concentration • Water diffuses across a membrane from the region of lower solute concentration to the region of higher solute concentration 7 9/15/2014 Water Balance of Cells Without Walls • Tonicity is the ability of a solution to cause a cell to gain -
Questions in Cell Biology
Name: Questions in Cell Biology Directions: The following questions are taken from previous IB Final Papers on the subject of cell biology. Answer all questions. This will serve as a study guide for the next quiz on Monday 11/21. 1. Outline the process of endocytosis. (Total 5 marks) 2. Draw a labelled diagram of the fluid mosaic model of the plasma membrane. (Total 5 marks) 3. The drawing below shows the structure of a virus. II I 10 nm (a) Identify structures labelled I and II. I: ...................................................................................................................................... II: ...................................................................................................................................... (2) (b) Use the scale bar to calculate the maximum diameter of the virus. Show your working. Answer: ..................................................... (2) (c) Explain briefly why antibiotics are effective against bacteria but not viruses. ............................................................................................................................................... ............................................................................................................................................... ............................................................................................................................................... .............................................................................................................................................. -
An Overview of Lipid Membrane Models for Biophysical Studies
biomimetics Review Mimicking the Mammalian Plasma Membrane: An Overview of Lipid Membrane Models for Biophysical Studies Alessandra Luchini 1 and Giuseppe Vitiello 2,3,* 1 Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, 2100 Copenhagen, Denmark; [email protected] 2 Department of Chemical, Materials and Production Engineering, University of Naples Federico II, Piazzale Tecchio 80, 80125 Naples, Italy 3 CSGI-Center for Colloid and Surface Science, via della Lastruccia 3, 50019 Sesto Fiorentino (Florence), Italy * Correspondence: [email protected] Abstract: Cell membranes are very complex biological systems including a large variety of lipids and proteins. Therefore, they are difficult to extract and directly investigate with biophysical methods. For many decades, the characterization of simpler biomimetic lipid membranes, which contain only a few lipid species, provided important physico-chemical information on the most abundant lipid species in cell membranes. These studies described physical and chemical properties that are most likely similar to those of real cell membranes. Indeed, biomimetic lipid membranes can be easily prepared in the lab and are compatible with multiple biophysical techniques. Lipid phase transitions, the bilayer structure, the impact of cholesterol on the structure and dynamics of lipid bilayers, and the selective recognition of target lipids by proteins, peptides, and drugs are all examples of the detailed information about cell membranes obtained by the investigation of biomimetic lipid membranes. This review focuses specifically on the advances that were achieved during the last decade in the field of biomimetic lipid membranes mimicking the mammalian plasma membrane. In particular, we provide a description of the most common types of lipid membrane models used for biophysical characterization, i.e., lipid membranes in solution and on surfaces, as well as recent examples of their Citation: Luchini, A.; Vitiello, G. -
Membrane Proteins Are Associated with the Membrane of a Cell Or Particular Organelle and Are Generally More Problematic to Purify Than Water-Soluble Proteins
Strategies for the Purification of Membrane Proteins Sinéad Marian Smith Department of Clinical Medicine, School of Medicine, Trinity College Dublin, Ireland. Email: [email protected] Abstract Although membrane proteins account for approximately 30 % of the coding regions of all sequenced genomes and play crucial roles in many fundamental cell processes, there are relatively few membranes with known 3D structure. This is likely due to technical challenges associated with membrane protein extraction, solubilization and purification. Membrane proteins are classified based on the level of interaction with membrane lipid bilayers, with peripheral membrane proteins associating non- covalently with the membrane, and integral membrane proteins associating more strongly by means of hydrophobic interactions. Generally speaking, peripheral membrane proteins can be purified by milder techniques than integral membrane proteins, whose extraction require phospholipid bilayer disruption by detergents. Here, important criteria for strategies of membrane protein purification are addressed, with a focus on the initial stages of membrane protein solublilization, where problems are most frequently are encountered. Protocols are outlined for the successful extraction of peripheral membrane proteins, solubilization of integral membrane proteins, and detergent removal which is important not only for retaining native protein stability and biological functions, but also for the efficiency of downstream purification techniques. Key Words: peripheral membrane protein, integral membrane protein, detergent, protein purification, protein solubilization. 1. Introduction Membrane proteins are associated with the membrane of a cell or particular organelle and are generally more problematic to purify than water-soluble proteins. Membrane proteins represent approximately 30 % of the open-reading frames of an organism’s genome (1-4), and play crucial roles in basic cell functions including signal transduction, energy production, nutrient uptake and cell-cell communication. -
Is Lipid Translocation Involved During Endo- and Exocytosis?
Biochimie 82 (2000) 497−509 © 2000 Société française de biochimie et biologie moléculaire / Éditions scientifiques et médicales Elsevier SAS. All rights reserved. S0300908400002091/FLA Is lipid translocation involved during endo- and exocytosis? Philippe F. Devaux* Institut de Biologie Physico-Chimique, UPR-CNRS 9052, 13, rue Pierre-et-Marie-Curie, 75005 Paris, France (Received 28 January 2000; accepted 17 March 2000) Abstract — Stimulation of the aminophospholipid translocase, responsible for the transport of phosphatidylserine and phosphati- dylethanolamine from the outer to the inner leaflet of the plasma membrane, provokes endocytic-like vesicles in erythrocytes and stimulates endocytosis in K562 cells. In this article arguments are given which support the idea that the active transport of lipids could be the driving force involved in membrane folding during the early step of endocytosis. The model is sustained by experiments on shape changes of pure lipid vesicles triggered by a change in the proportion of inner and outer lipids. It is shown that the formation of microvesicles with a diameter of 100–200 nm caused by the translocation of plasma membrane lipids implies a surface tension in the whole membrane. It is likely that cytoskeleton proteins and inner organelles prevent a real cell from undergoing overall shape changes of the type seen with giant unilamellar vesicles. Another hypothesis put forward in this article is the possible implication of the phospholipid ‘scramblase’ during exocytosis which could favor the unfolding of microvesicles. © 2000 Société française de biochimie et biologie moléculaire / Éditions scientifiques et médicales Elsevier SAS aminophospholipid translocase / membrane budding / spontaneous curvature / liposomes / K562 cells 1. Introduction yet whether clathrin polymerizes and then pinches off the membrane to form the buds or if polymerization takes During the last 10–15 years, a large number of proteins place around a pre-formed bud. -
The Membrane
The Membrane Natalie Gugala1*, Stephana J Cherak1 and Raymond J Turner1 1Department of Biological Sciences, University of Calgary, Canada *Corresponding author: RJ Turner, Department of Biological Sciences, University of Calgary, Alberta, Canada, Tel: 1-403-220-4308; Fax: 1-403-289-9311; Email: [email protected] Published Date: February 10, 2016 ABSTRACT and continues to be studied. The biological membrane is comprised of numerous amphiphilic The characterization of the cell membrane has significantly extended over the past century lipids, sterols, proteins, carbohydrates, ions and water molecules that result in two asymmetric polar leaflets, in which the interior is hydrophobic due to the hydrocarbon tails of the lipids. generated a dynamic heterogonous image of the membrane that includes lateral domains and The extension of the Fluid Mosaic Model, first proposed by Singer and Nicolson in 1972, has clusters perpetrated by lipid-lipid, protein-lipid and protein-protein interactions. Proteins found within the membrane, which are generally characterized as either intrinsic or extrinsic, have an array of biological functions vital for cell activity. The primary role of the membrane, among many, is to provide a barrier that conveys both separation and protection, thus maintaining the integrity of the cell. However, depending on the permeability of the membrane several ions are able to move down their concentration gradients. In turn this generates a membrane potential difference between the cytosol, which is found to have an excess negative charge, and surrounding extracellular fluid. Across a biological cell membrane, several potentials can be found. These include the Nernst or equilibrium potential, in which there is no overall flow of a Basicparticular Biochemistry ion and | www.austinpublishinggroup.com/ebooks the Donnan potential, created by an unequal distribution of ions. -
Biological Membranes
14 Biological Membranes To understand the structure The fundamental unit of life is the cell. All living things are composed of Goal and composition of biological cells, be it a single cell in the case of many microorganisms or a highly membranes. organized ensemble of myriad cell types in the case of multicellular organisms. A defining feature of the cell is a membrane, the cytoplasmic Objectives membrane, that surrounds the cell and separates the inside of the cell, the After this chapter, you should be able to cytoplasm, from other cells and the extracellular milieu. Membranes also • distinguish between cis and trans surround specialized compartments inside of cells known as organelles. unsaturated fatty acids. Whereas cells are typically several microns (μm) in diameter (although • explain why phospholipids some cells can be much larger), the membrane is only about 10 nanometers spontaneously form lipid bilayers and (nm) thick. Yet, and as we will see in subsequent chapters, the membrane is sealed compartments. not simply an ultra-thin, pliable sheet that encases the cytoplasm. Rather, • describe membrane fluidity and how it membranes are dynamic structures that mediate many functions in the is affected by membrane composition life of the cell. In this chapter we examine the composition of membranes, and temperature. their assembly, the forces that stabilize them, and the chemical and physical • explain the role of cholesterol in properties that influence their function. buffering membrane fluidity. The preceding chapters have focused on two kinds of biological molecules, • explain how the polar backbone namely proteins and nucleic acids, that are important in the workings of a membrane protein can be accommodated in a bilayer. -
The Lipid Bilayer: Composition and Structural Organization
THE LIPID BILAYER: COMPOSITION AND STRUCTURAL ORGANIZATION • MR. SOURAV BARAI • ASSISTANT PROFESSOR • DEPARTMENT OF ZOOLOGY • JHARGRAM RAJ COLLEGE THELIPID BILAYER: COMPOSITION AND STRUCTURAL ORGANIZATION The Fluid Mosaic Model of Biomembrane Plasma membrane • 1. Affect shape and function • 2. Anchor protein to the membrane • 3. Modify membrane protein activities • 4. Transducing signals to the cytoplasm “A living cell is a self-reproducing system of molecules held inside a container - the plasma membrane” Membrane comprised of lipid sheet (5 nm thick) • Primary purpose - barrier to prevent cell contents spilling out BUT, must be selective barrier Lipid Composition and struCturaL organization • Phospholipids of the composition present in cells spontaneously form sheet like phospholipid bilayers, which are two molecules thick. • The hydrocarbon chains of the phospholipids in each layer, or leaflet, form a hydrophobic core that is 3–4 nm thick in most biomembranes. • Approx 10^6 lipid molecule in 1µm×1µm area of lipid bilayer. • Electron microscopy of thin membrane sections stained with osmium tetroxide, which binds strongly to the polar head groups of phospholipids, reveals the bilayer structure. • A cross section of all single membranes stained with osmium tetroxide looks like a railroad track: two thin dark lines (the stain–head group complexes) with a uniform light space of about 2nm (the hydrophobic tails) between them. PROPERTIES • PERMIABILITY: The hydrophobic core is an impermeable barrier that prevents the diffusion of water-soluble (hydrophilic) solutes across the membrane. • STABILITY: The bilayer structure is maintained by hydrophobic and van der Waals interactions between the lipid chains. Even though the exterior aqueous environment can vary widely in ionic strength and pH, the bilayer has the strength to retain its characteristic architecture.