2009 Lecture 3

From CellBiology
Eukaryotic Cell Physical Compartments

Cell Membranes and Compartments

Introduction

A major difference between eukayotes and prokaryotes is the presence of physical compartments (membrane bound) within the cell. These compartments allow the separation/specialization of processes within the cell. There also exist within each of these physical compartments, functional compartments where specific processes may occur or are restricted too. This lecture is an introduction to compartments within the cell and membranes. The key components are: cell compartments, membrane structure, membrane models, membrane specializations.

Dr Mark Hill (2009)

Lecture Audio

The University has a system for automated recording of lectures called Lectopia. Lectopia requires login using your student number and unipass. I will be adding the link to each iLecture Audio following the Lecture. Due to the automated recording method, most lectures begin 4-5 minutes into MP3 recordings and occasionally stop before the end of the lecture. 2009 All Audio Files

Lecture 3: Cell Membranes and Compartments Lecture Date: 17-03-2009 Lecture Time: 13:00 Venue: BioMed F Speaker: Mark Hill

Archive

MH - note that content listed below will not match exactly current lecture structure but has been selected as having similar content.

Objectives

  • Understand the concept of separate intracellular spaces
  • Understand the structure of membranes
  • Brief understanding of history of membrane models
  • Understand the difference between physical and functional compartments
  • Brief understanding of membrane specializations

History

Cork Bark by Robert Hooke 1665
  • Robert Hooke (1635-1703)
    • used early microscopes to view cork tree bark
    • was the first to use the term CELL
  • Robert Brown 1825
    • identified nuclei in plant cells
  • Theodor Schwann (1810 - 1882)
    • together with Matthias Schleiden (plants) developed the cell theory in 1839

Cell Theory

    • All organisms consist of one or more cells
    • The cell is the basic unit of structure for all cells
    • All cells arise only from preexisting cells

Plasma Membrane Images

Encloses or covers or cell types. Some different ways of looking microscopically at membranes.

Membrane - Light Micrograph

CellB35x10L2.jpg

Eo104le.jpg

Membrane - Fluorescent Micrograph

Candida albicans.jpg

Yeast - Candida albicans

Caveolae.gif

Lipid raft microdomain compartmentalization of TC10 is required for insulin signaling and GLUT4 translocation. Watson RT, Shigematsu S, Chiang SH, Mora S, Kanzaki M, Macara IG, Saltiel AR, Pessin JE. J Cell Biol. 2001 Aug 20;154(4):829-40. Epub 2001 Aug 13. PMID: 11502760

Membrane - Scanning Electron Micrograph

Red White Blood cells.jpg

Mycobacterium tuberculosis bacteria.jpg


Membrane - Transmission Electron Micrograph

Plasma and Organelle Membranes


CDCcandida.jpg

Yeast - Candida albicans


Links: Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure Denk W, Horstmann H (2004) Serial Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure. PLoS Biol 2(11): e329

Compartments

  • Physical Compartments
    • membrane bound
    • Nucleus, Cytoplasm, Organelles
    • cell nomenclature based upon presence or absence of these compartments (eukaryotic, prokaryotic)
  • Functional Compartments
    • spatial localization
    • targeting
    • activation and inactivation
    • signaling

Major Cellular Compartments

Proposed model for organelle membrane evolution
  • Nucleus (nuclear) - contains a single organelle compartment
  • Cytoplasm (cytoplasmic) - contains many organelle compartments

Organelle Number/Volume

  • How many organelles?
  • How much space within the cell do they occupy?
  • Are all the cells the same?

Take a typical mammalian liver cell....


Liver Structure


Table 12-1. Relative Volumes Occupied by the Major Intracellular Compartments in a Liver Cell (Hepatocyte)

Table 12-2. Relative Amounts of Membrane Types in Two Kinds of Eucaryotic Cells

Compartments are Dynamic

Movies showing flexibility of membranes and their changing shape and size.


Nuclear Compartment

  • Nuclear matrix - consisting of Intermediate filaments (lamins)
  • Nucleoli (functional compartment - localised transcription DNA of RNA genes)
  • Chromosomes (DNA and associated proteins)

(MH - you will not see chromosomes in interphase nuclei only during mitosis, more in the Nucleus Lecture)

Cytoplasmic Compartment

  • Cytoplasmic Organelles
    • Membrane bound structures
    • Endoplasmic reticulum, golgi apparatus, mitochondria, lysosomes, peroxisomes, vesicles
  • Cytoskeleton
    • 3 filament systems
  • Cytoplasmic “structures”
    • Ribosomes
    • DNA -> mRNA -> Protein
    • Proteins
    • Receptors, signaling, metabolism, structural
    • Viruses, bacteria, prionsl
  • Functional compartments
    • occur in nucleus, cytoplasm, in organelles and outside organelles
    • signaling, metabolic reactions, processing genetic information, cytoskeleton dynamics, vesicle dynamics

Membrane Functions

  • Form compartments
  • Allow “specialization”
  • Metabolic and biochemical
  • Localization of function
  • Regulation of transport
  • Detection of signals
  • Cell-cell communication
  • Cell Identity

Plasma Membrane Image

EM - Cell (Plasma) and Organelle Membranes

Cell (Plasma) encloses or covers or cell types.

Tem cellstructures2.jpg

Cell (Plasma) and Organelle Membranes

Membrane Components

Model of Cell (plasma) membrane structure
  • phospholipids, proteins and cholesterol
  • first compartment formed
  • prokaryotes (bacteria) just this 1 compartment
  • eukaryotic cells many different compartments

Phospholipids

  • membranes contain phospholipids, glycolipids, and steroids
  • The main lipid components include:
    • phosphatidylcholine (~50%)
    • phosphatidylethanolamine (~10%)
    • phosphatidylserine (~15%)
    • sphingolipids (~10%)
    • cholesterol (~10%)
    • phosphatidylinositol (1%).


Links: Three views of a cell membrane

Phospholipid Orientation

Figure 2-22. Phospholipid structure and the orientation of phospholipids in membranes


Phospholipid Bilayer
  • A liposome (lipid vesicle) is a small aqueous compartment surrounded by a lipid bilayer.
  • A micelle is a small compartment surrounded by a single lipid layer.
Phospholipid Orientation

Membranes History

  • 1890 Charles Overton
    • selective permeation of membranes
    • non-polar pass through (lipid soluble)
    • polar refractory
    • lipids present as a coat
  • 1905 Irving Langmuir
    • lipids faced with heads towards water away from organic solvents
  • 1925 Gorter and Grendel
    • monolayer of lipid isolated from rbc
    • twice (2x) surface area of cell (bilayer)
  • 1930-40 Danielle-Davson
    • Proteins coat a bilayer with polar “pores”
  • 1960s Robertson
    • Modification with glycoprotein on one side, therefore asymmetric
  • 1972 Singer and Nicholson
    • proteins “floating” within lipid bilayer like a “liquid” surface
  • 1975 Unwin and Henderson
    • integral membrane proteins
    • both hydrophobic and hydrophilic
    • alternating -phobic and -philic represent trans-membrane loops
    • glycoprotein carbohydrate groups on outer surface

Membranes Recent History

Lipid rafts
  • 1997 Simons
    • cholesterol to form rafts that move within the fluid bilayer
    • “Membrane Rafts”
    • “A new aspect of cell membrane structure is presented, based on the dynamic clustering of sphingolipids and cholesterol to form rafts that move within the fluid bilayer. It is proposed that these rafts function as platforms for the attachment of proteins when membranes are moved around inside the cell and during signal transduction.”

Reference: Simons K, Ikonen E. Nature 1997 Jun 5;387(6633):569-72

Membrane Proteins

  • Proteins can be embedded in the inner phospholipid layer, outer phospholipid layer or span both layers
  • Some proteins are folded such that they span the membrane in a series of “loops”

Membrane Protein Functions

  • transport channels
  • enzyme reactions
  • cytoskeleton link
  • cell adhesion
  • cell identity
Links: Figure 17-21. Topologies of some integral membrane proteins synthesized on the rough ER

Membrane Glycoproteins

  • Glycoproteins are proteins which have carbohydrate groups (sugars) attached
  • to produce these proteins go through a very specific cellular pathway of organelles (secretory pathway)
  • to reach the cell surface where they are either secreted (form part of the extracellular matrix)
  • or are embedded in the membrane with the carbohydrate grouped on the outside surface (integral membrane protein)

Membrane Cholesterol

Model of Cell (plasma) membrane structure
  • Small molecule embedded between the phospholipid molecules and regulates lipid mobility (MH - see rafts)
  • Cholesterol can be at different concentrations in different regions of plasma membrane
  • lateral organization of membranes and free volume distribution
  • may control membrane protein activity and "raft” formation
  • fine tuning of membrane lipid composition, organization/dynamics, function
  • bacterial membranes (except for Mycoplasma and some methylotrophic bacteria) have no sterols, they lack the enzymes required for sterol biosynthesis.
Links:  MBoC Figure 10-9. Cholesterol in a lipid bilayer

Bacterial Membranes

(MH - covered this in previous Lecture)

  • Bacteria with double membranes (Example: E. coli)
    • inner membrane is the cell's plasma membrane
    • Gram Negative do not retain dark blue dye used in gram staining
  • Bacteria with single membranes (Example: staphylo-cocci and streptococci)
    • thicker cell walls
    • Gram Positive because they do retain blue dye
    • single membrane comparable to inner (plasma) membrane of gram negative bacteria


Membrane Fluidity

  • fusion of 2 cells
  • FRAP
  • membrane domains (polarized cells)
    • epithelia - apical, basal and lateral domains
Links: FRAP |  MBC - Membrane Fluidity

Membrane Specializations

  • plasma membrane cytoskeleton
  • different directly under membranes
  • adhesion complexes
  • absorbtive and secretory
  • synaptic junctions
This range of membrane specializations will be covered in detail in later Lectures.

Adhesion Specializations

A series of different types of proteins and cytoskeleton associations forming different classes of adhesion junctions (MH - covered in detail in a lecture 8)

  • Desmosomes ( = macula adherens)
  • Adherens Junctions ( = zonula adherens)
  • Septate Junctions
  • Tight Junctions
  • Gap Junctions
Covered in  Lecture 8 - Cell Junctions

Membrane Transport

Three major forms of transport across the membrane

  • Passive - Simple diffusion
  • Facilitated - transport proteins
  • Active - transport proteins for nutrient uptake, secretion, ion balance

Ion Channels

  • phospholipid impermeable to ions in aqueous solution
  • protein channels permit rapid ion flux
  • 1960’s structure and function, ionophores (simple ion channels)
  • common structural motif alpha helix
  • 75 + different ion channels
  • Allosteric proteins - conformation regulated by different stimuli
  • opening/closing, “gating” of ions
Cell potassium channels

Ion Channel Types

  • 3 rapid + 1 slow gate (gap junction)
    • Voltage-gated - propogation of electrical signals along nerve, muscle
    • Ligand-gated - opened by non-covalent, reversible binding of ligand between nerve cells, nerve-muscle, gland cells
    • Mechanical-gated - regulated by mechanical deformation
    • Gap junction - allow ions to flow between adjacent cells open/close in response to Ca2+ and protons

Membrane Transport Disease

  • Cystic Fibrosis
    • 1989 Collins (US), Tsui and Riordan (Canada)
    • Chloride channel protein mutation
    • point mutant, folded improperly, trapped and degraded in ER

Historic Papers

Below are some example historical research finding related to cell membranes from the JCB Archive and other sources.

1957 The invention of freeze fracture EM and the determination of membrane structure Russell Steere introduces his home-made contraption for freeze fracture electron microscopy (EM), and Daniel Branton uses it to conclude that membranes are bilayers.

1971 Spectrin is peripheral S. Jonathan Singer, Garth Nicolson, and Vincent Marchesi use red cell ghosts to provide strong evidence for the existence of peripheral membrane proteins.

1992 Lipid raft idea is floated Gerrit van Meer and Kai Simons get the first hints of lipid rafts based on lipid sorting experiments.


Links:  Sorted JCB Archive -Membranes JCB Archive

References

Textbooks

Search Online Textbooks

Books

Reviews

Vertebrate membrane proteins: structure, function, and insights from biophysical approaches. Müller DJ, Wu N, Palczewski K. Pharmacol Rev. 2008 Mar;60(1):43-78. Epub 2008 Mar 5. Review. PMID: 18321962

Articles

Images

Proposed model for organelle membrane evolution
Model of Cell (plasma) membrane structure
Model of Exocytosis mechanisms also showing new membrane turnover.




Page References

Membrane Terms

  • adhesion junction membrane specialization allowing either cell-cell or cell-extracellular matrix adhesion generally in multicellular organisms.
  • bilayer having two layers, refers to the 2 lipid layers of a single membrane.
  • blebbing a plasma membrane change often associated with apoptosis. The underlying cell cytoskeleton is disrupted leading to a the bubbling of the plasma membrane, which will enclose cytoplasmic and nuclear components.
  • cholesterol bacterial membranes (except for Mycoplasma and some methylotrophic bacteria) have no sterols, they lack the enzymes required for sterol biosynthesis.
  • electron microscopy a microscope technique that uses beams of electrons instead of light to generate high resolution images of cellular components. This technique historically gave the first images of the membrane bilayer structure.
  • electron tomography an electron microscopic technique to generate a three dimensional (3-D) image from any electron microscopy specimen.
  • flippases enzymes that catalyze rapid translocation of phospholipids across the endoplasmic reticulum membrane. Required for balanced growth of both halves of the bilayer.
  • functional compartment a specialized region formed within a cell which is not limited by a membrane, compared to a "structural compartment".
  • Gram negative term used to describe bacteria which do not retain the Gram dye when stained. These are bacteria with double membranes, the inner membrane is the cell's plasma membrane (Example: E. coli).
  • Gram positive term used to describe bacteria which do retain the Gram dye when stained. These are bacteria with single membranes and thicker cell wall (Example: staphylo-cocci and streptococci).
  • lipids the basic molecules forming the lipid bilayer as phospholipids, glycolipids, and steroids. The main lipid components include phosphatidylcholine (~50%), phosphatidylethanolamine (~10%), phosphatidylserine (~15%), sphingolipids (~10%), cholesterol (~10%), and phosphatidylinositol (1%). Medical Microbiology - Plasma (Cytoplasmic) Membranes
  • liposome (lipid vesicle) is a small aqueous compartment surrounded by a lipid bilayer.
  • membrane cytoskeleton the components of the cell cytoskeleton that directly underly either the cell (plasma) and nuclear membranes.
  • micelle is a small compartment surrounded by a single lipid layer.
  • phospholipid the basic molecule forming the lipid bilayer of a typical membrane (see also lipid).
  • structural compartment a specialized region formed within a cell which is limited by a membrane, compared to a "functional compartment".
  • raft (lipid rafts) term used to describe stabilized regions that form within membranes. These rafts "float" within the lipid membrane and are formed by cholesterol altering (stabilizing) the fluidity of the local membrane.
  • vesicle general term given to any membrane enclosing material within the cytoplasm.
  • protein-to-lipid ratio the analysis of membranes by separating the 2 main components. For example, bacterial plasma membranes are approximately 3:1, close to those for mitochondrial membranes.

Internal Links

External Links

Working Area

The following material is a draft notebook area, not part of the current lecture content.

  • Vertebrate membrane proteins: structure, function, and insights from biophysical approaches. Müller DJ, Wu N, Palczewski K. Pharmacol Rev. 2008 Mar;60(1):43-78. Epub 2008 Mar 5. Review. PMID: 18321962
  • Cell motility through plasma membrane blebbing Oliver T. Fackler and Robert Grosse J Cell Biol. 2008 June 16; 181(6): 879–884. doi: 10.1083/jcb.200802081. PMCID: PMC2426937
  • Three-dimensional reconstruction of the membrane skeleton at the plasma membrane interface by electron tomography. Morone N, Fujiwara T, Murase K, Kasai RS, Ike H, Yuasa S, Usukura J, Kusumi A. J Cell Biol. 2006 Sep 11;174(6):851-62. Epub 2006 Sep 5. PMID: 1695434 JCB http://jcb.rupress.org/cgi/content/full/174/6/851?


2009 Course Content

Lectures

Cell Biology Introduction | Cells Eukaryotes and Prokaryotes | Cell Membranes and Compartments | Cell Nucleus | Cell Export - Exocytosis | Cell Import - Endocytosis | Cell Mitochondria | Cell Junctions | Cytoskeleton Introduction | Cytoskeleton 1 Intermediate Filaments | Cytoskeleton 2 Microtubules | Cytoskeleton 3 Microfilaments | Extracellular Matrix 1 | Extracellular Matrix 2 | Cell Cycle | Cell Division | Cell Death 1 | Cell Death 2 | Signal 1 | Signal 2 | Stem Cells | Stem Cells | Development | Revision

Laboratories

Introduction to Lab | Microscopy Methods | Preparation/Fixation | Immunochemistry | Cell Knockout Methods | Cytoskeleton Exercise | Confocal Microscopy | Tissue Culture 1 | Tissue Culture 2 | Microarray Lab visit

Dr Mark Hill 2015, UNSW Cell Biology - UNSW CRICOS Provider Code No. 00098G