Structure and Activity of Lipid Bilayer Within a Membrane-Protein Transporter
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Spring 2013 Lecture 23
CHM333 LECTURES 23: 3/25/13 SPRING 2013 Professor Christine Hrycyna LIPIDS III EFFECT OF CHOLESTEROL ON MEMBRANES: - Bulky rigid molecule - Moderates fluidity of membranes – both increases and decreases o Cholesterol in membranes DECREASES fluidity because it is rigid o Prevents crystallization (making solid) of fatty acyl side chains by fitting between them. Disrupts close packing of fatty acyl chains. Therefore, INCREASED fluidity BIOLOGICAL MEMBRANES CONTAIN PROTEINS AS WELL AS LIPIDS: - Proteins are 20-80% of cell membrane - Rest is lipid or carbohydrate; supramolecular assembly of lipid, protein and carbohydrate - Proteins are also distributed asymmetrically - TWO classes of Membrane Proteins: o Integral Membrane Proteins o Peripheral Membrane Proteins 178 CHM333 LECTURES 23: 3/25/13 SPRING 2013 Professor Christine Hrycyna - INTEGRAL MEMBRANE PROTEINS o Located WITHIN the lipid bilayer o Usually span the bilayer one or more times – called transmembrane (TM) proteins o Hydrophobic amino acids interact with fatty acid chains in the hydrophobic core of the membrane o Can be removed from the membrane with detergents like SDS – need to disrupt the hydrophobic interactions § Membrane Disruption Animation: o http://www.youtube.com/watch?v=AHT37pvcjc0 o Function: § Transporters – moving molecules into or out of cells or cell membranes § Receptors – transmitting signals from outside of the cell to the inside - β Barrel Integral Membrane Proteins § Barrel-shaped membrane protein that is made up of antiparallel β-strands with hydrophilic (interior) and hydrophobic (facing lipid tails). § So far found only in outer membranes of Gram-negative bacteria, cell wall of Gram-positive bacteria, and outer membranes of mitochondria and chloroplasts. 179 CHM333 LECTURES 23: 3/25/13 SPRING 2013 Professor Christine Hrycyna - α-Helical Membrane Proteins - Can cross the membrane once or many times and have multiple transmembrane segments. -
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. -
Predicting Protein-Membrane Interfaces of Peripheral Membrane
bioRxiv preprint doi: https://doi.org/10.1101/2021.06.28.450157; this version posted June 29, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Predicting protein-membrane interfaces of pe- ripheral membrane proteins using ensemble machine learning Alexios Chatzigoulas1,2,* and Zoe Cournia1,* 1Biomedical Research Foundation, Academy of Athens, 4 Soranou Ephessiou, 11527 Athens, Greece, 2Depart- ment of Informatics and Telecommunications, National and Kapodistrian University of Athens, 15784 Athens, Greece *To whom correspondence should be addressed. Abstract Motivation: Abnormal protein-membrane attachment is involved in deregulated cellular pathways and in disease. Therefore, the possibility to modulate protein-membrane interactions represents a new promising therapeutic strategy for peripheral membrane proteins that have been considered so far undruggable. A major obstacle in this drug design strategy is that the membrane binding domains of peripheral membrane proteins are usually not known. The development of fast and efficient algorithms predicting the protein-membrane interface would shed light into the accessibility of membrane-protein interfaces by drug-like molecules. Results: Herein, we describe an ensemble machine learning methodology and algorithm for predicting membrane-penetrating residues. We utilize available experimental data in the literature for training 21 machine learning classifiers and a voting classifier. Evaluation of the ensemble classifier accuracy pro- duced a macro-averaged F1 score = 0.92 and an MCC = 0.84 for predicting correctly membrane-pen- etrating residues on unknown proteins of an independent test set. -
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. -
The Structure of Biological Membranes
The Structure of Biological Membranes Func7ons of Cellular Membranes 1. Plasma membrane acts as a selecvely permeable barrier to the environment • Uptake of nutrients • Waste disposal • Maintains intracellular ionic milieau 2. Plasma membrane facilitates communicaon • With the environment • With other cells • Protein secreon 3. Intracellular membranes allow compartmentalizaon and separaon of different chemical reac7on pathways • Increased efficiency through proximity • Prevent fu8le cycling through separaon Composi7on of Animal Cell Membranes • Hydrated, proteinaceous lipid bilayers • By weight: 20% water, 80% solids • Solids: Lipid Protein ~90% Carbohydrate (~10%) • Phospholipids responsible for basic membrane bilayer structure and physical proper8es • Membranes are 2-dimensional fluids into which proteins are dissolved or embedded The Most Common Class of PhospholipiD is FormeD from a Gycerol-3-P Backbone SaturateD FaJy AciD •Palmitate and stearate most common •14-26 carbons •Even # of carbons UnsaturateD FaJy AciD Structure of PhosphoglyceriDes All Membrane LipiDs are Amphipathic Figure 10-2 Molecular Biology of the Cell (© Garland Science 2008) PhosphoglyceriDes are Classified by their Head Groups Phosphadylethanolamine Phosphadylcholine Phosphadylserine Phosphadylinositol Ether Bond at C1 PS and PI bear a net negave charge at neutral pH Sphingolipids are the SeconD Major Class of PhospholipiD in Animal Cells Sphingosine Ceramides contain sugar moies in ether linkage to sphingosine GlycolipiDs are AbunDant in Brain Cells Figure 10-18 Molecular -
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 Membrane Interface of Chloroplast Signal Recognition Particle-Dependent Protein Targeting
University of Arkansas, Fayetteville ScholarWorks@UARK Theses and Dissertations 5-2009 The eM mbrane Interface of Chloroplast Signal Recognition Particle-Dependent Protein Targeting Naomi Jane Marty University of Arkansas, Fayetteville Follow this and additional works at: http://scholarworks.uark.edu/etd Part of the Cell Biology Commons, and the Molecular Biology Commons Recommended Citation Marty, Naomi Jane, "The eM mbrane Interface of Chloroplast Signal Recognition Particle-Dependent Protein Targeting" (2009). Theses and Dissertations. 6. http://scholarworks.uark.edu/etd/6 This Dissertation is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of ScholarWorks@UARK. For more information, please contact [email protected], [email protected]. THE MEMBRANE INTERFACE OF CHLOROPLAST SIGNAL RECOGNITION PARTICLE-DEPENDENT PROTEIN TARGETING THE MEMBRANE INTERFACE OF CHLOROPLAST SIGNAL RECOGNITION PARTICLE-DEPENDENT PROTEIN TARGETING A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Cell and Molecular Biology By Naomi J. Marty Northeastern State University Bachelor of Science in Biology, 2003 May 2009 University of Arkansas ABSTRACT A novel signal recognition particle (SRP) found in the chloroplast (cpSRP) works in combination with the cpSRP receptor, cpFtsY, to facilitate the post-translational targeting of a family of nuclear-encoded thylakoid proteins to the Alb3 translocase in thylakoid membranes. Work here focused on understanding events at the membrane that take place to ensure targeting of the cpSRP-dependent substrate to Alb3. Specifically, we sought to understand the structural and functional role of membrane binding by cpFtsY, a protein that exhibits the ability to partition between the membrane (thylakoid) and soluble (stroma) phase during protein targeting. -
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. -
Therapeutic Nanobodies Targeting Cell Plasma Membrane Transport Proteins: a High-Risk/High-Gain Endeavor
biomolecules Review Therapeutic Nanobodies Targeting Cell Plasma Membrane Transport Proteins: A High-Risk/High-Gain Endeavor Raf Van Campenhout 1 , Serge Muyldermans 2 , Mathieu Vinken 1,†, Nick Devoogdt 3,† and Timo W.M. De Groof 3,*,† 1 Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium; [email protected] (R.V.C.); [email protected] (M.V.) 2 Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium; [email protected] 3 In Vivo Cellular and Molecular Imaging Laboratory, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium; [email protected] * Correspondence: [email protected]; Tel.: +32-2-6291980 † These authors share equal seniorship. Abstract: Cell plasma membrane proteins are considered as gatekeepers of the cell and play a major role in regulating various processes. Transport proteins constitute a subclass of cell plasma membrane proteins enabling the exchange of molecules and ions between the extracellular environment and the cytosol. A plethora of human pathologies are associated with the altered expression or dysfunction of cell plasma membrane transport proteins, making them interesting therapeutic drug targets. However, the search for therapeutics is challenging, since many drug candidates targeting cell plasma membrane proteins fail in (pre)clinical testing due to inadequate selectivity, specificity, potency or stability. These latter characteristics are met by nanobodies, which potentially renders them eligible therapeutics targeting cell plasma membrane proteins. Therefore, a therapeutic nanobody-based strategy seems a valid approach to target and modulate the activity of cell plasma membrane Citation: Van Campenhout, R.; transport proteins. -
S41467-021-23799-1.Pdf
ARTICLE https://doi.org/10.1038/s41467-021-23799-1 OPEN Nanoscale architecture of a VAP-A-OSBP tethering complex at membrane contact sites ✉ Eugenio de la Mora1,2,5, Manuela Dezi 1,2,5 , Aurélie Di Cicco1,2, Joëlle Bigay 3, Romain Gautier 3, ✉ John Manzi 1,2, Joël Polidori3, Daniel Castaño-Díez4, Bruno Mesmin 3, Bruno Antonny 3 & ✉ Daniel Lévy 1,2 Membrane contact sites (MCS) are subcellular regions where two organelles appose their 1234567890():,; membranes to exchange small molecules, including lipids. Structural information on how proteins form MCS is scarce. We designed an in vitro MCS with two membranes and a pair of tethering proteins suitable for cryo-tomography analysis. It includes VAP-A, an ER trans- membrane protein interacting with a myriad of cytosolic proteins, and oxysterol-binding protein (OSBP), a lipid transfer protein that transports cholesterol from the ER to the trans Golgi network. We show that VAP-A is a highly flexible protein, allowing formation of MCS of variable intermembrane distance. The tethering part of OSBP contains a central, dimeric, and helical T-shape region. We propose that the molecular flexibility of VAP-A enables the recruitment of partners of different sizes within MCS of adjustable thickness, whereas the T geometry of the OSBP dimer facilitates the movement of the two lipid-transfer domains between membranes. 1 Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, CNRS UMR168, Paris, France. 2 Sorbonne Université, Paris, France. 3 CNRS UMR 7275, Université Côte d’Azur, Institut de Pharmacologie Moléculaire et Cellulaire, Valbonne, France. 4 BioEM Lab, C-CINA, Biozentrum, University of Basel, ✉ Basel, Switzerland.