Chapter 6: A Tour of the Cell Key Concepts 6.1 To study cells, biologists use microscopes and the tools of biochemistry. 6.2 Eukaryotic cells have internal membranes that compartmentalize their functions, and membrane bound organelles. 6.3 The Eukaryotic cell’s genetic instructions are found in the nucleus and carried out by the ribosomes. 6.4 The endomembrane system (ER) regulates protein traffic and performs metabolic function of the cell. 6.5 Mitochondria and chloroplasts change energy from one form to another. 6.6 The cytoskeleton is a network of fibers that organizes structures and activities inside the cell. 6.7 Extracellular components (outside the cell) and connections between cells (desmosomes, gap junctions, tight junctions, and plasmodesmata) help coordinate cellular activities. Microscopy • The study of cells progressed with the invention of the microscope in 1590, and their improvement in the 17th century. • Light Microscope- earliest form, uses beam of light passed thru specimen and then thru glass lenses. • Electron Microscope- (1950’s) focuses a beam of electrons thru a specimen, or onto its surface. Highest magnification and resolution. • Transmission EM- (TEM) used to study the internal structures of a cell. • Scanning EM- (SEM) used to study the surface of a specimen. Light Microscope TEM SEM Fig. 6-4 TECHNIQUE (a) Scanning electron microscopy (SEM) RESULTS Cilia 1 µm (b) Transmission electron Longitudinal Cross section section of of cilium microscopy (TEM) 1 µm cilium Parameters in Microscopy • Magnificationthe ratio of an objects image size to its real size • Resolution- a measure of the clarity of an object Staining Technique • The addition of certain chemical pigments allows for increased contrast between cell structures. • Makes structures easier to distinguish and study. • Examples- bromthymol blue, methylene blue, crystal violet, safranin, malachite green, eosin Fig. 6-3ab TECHNIQUE RESULTS (a) Brightfield (unstained specimen) 50 µm (b) Brightfield (stained specimen) Cell Fractionation •The goal of cell fractionation is to take cells apart and separate major organelles from one another. •The instrument used is a Centrifuge. •The centrifuge spins test tubes holding mixtures of disrupted cells at high speeds. •Uses centrifugal force. •Enable scientists to prepare specific components of cells in bulk quantities to study their composition and structure. Fig. 6-5 TECHNIQUE Homogenization Tissue cells Homogenate 1,000 g (1,000 times the force of gravity) Differential centrifugation 10 min Supernatant poured into next tube 20,000 g 20 min Pellet rich in nuclei and cellular debris 80,000 g 60 min 150,000 g 3 hr Pellet rich in mitochondria (and chloroplasts if cells are from a plant) Pellet rich in “microsomes” (pieces of plasma membranes and cells’ internal membranes) Pellet rich in ribosomes Fig. 6-5b TECHNIQUE (cont.) 1,000 g (1,000 times the force of gravity) 10 min Supernatant poured into next tube 20,000 g 20 min 80,000 g 60 min Pellet rich in nuclei and cellular debris 150,000 g 3 hr Pellet rich in mitochondria (and chloroplasts if cells are from a plant) Pellet rich in “microsomes” (pieces of plasma membranes and cells’ internal membranes) Pellet rich in ribosomes Cells of Living Things Prokaryotic Usually single celled. Can form colonies. No nucleus or membrane-bound organelles. Genetic material localized (nucleoid) Ex. Bacteria Eukaryotic Kingdoms: Protista, Fungi, Plants, Animals. Nuclear membrane encloses DNA. Organelles that have membrane. Cell Size and Shape • Surface to Volume Ratio limits size of cells. Large cells require more raw materials. • V = cm3 S.A. = cm2 Restrictions on size and shape • Cells compartmentalize to increase SA/Vol, specialize rxn within, localize reactions where needed. Basic Aspects of Cell Structure and Function • Plasma membrane • Lipid bilayer • Proteins – Channels, transport, pumps, receptors • DNA-containing region • Cytoplasm Prokaryotic Cells •Highly disorganized •No membrane bound organelles •Have a nucleoid (like a nucleus) •Nucleoid contains genetic material (DNA) •Have a plasma membrane •Have cytosol inside the cell in which the organelles are found •Have ribosomes •Smaller than Eukaryotic cells •Examples- Bacteria, Archae Typical Prokaryote (Bacterial) Cell Fig. 6-6 Fimbriae Nucleoid Ribosomes Plasma membrane Bacterial chromosome Cell wall Capsule 0.5 µm (a) A typical rod-shaped bacterium Flagella (b) A thin section through the bacterium Bacillus coagulans (TEM) Eukaryotic Cells • • • • Have membrane bound organelles Are very organized Have a nucleus (DNA) Surrounded by either a cell wall or a plasma membrane • Examples- plants, animals, fungi, protists (amoeba, paramecium,euglena) Defining Structures of Eukaryotic Cells A Plant Cell An Animal Cell Plant vs. Animal Cell Fig. 6-9a Nuclear envelope ENDOPLASMIC RETICULUM (ER) Flagellum Rough ER NUCLEUS Nucleolus Smooth ER Chromatin Centrosome Plasma membrane CYTOSKELETON: Microfilaments Intermediate filaments Microtubules Ribosomes Microvilli Golgi apparatus Peroxisome Mitochondrion Lysosome Fig. 6-9b NUCLEUS Nuclear envelope Nucleolus Chromatin Rough endoplasmic reticulum Smooth endoplasmic reticulum Ribosomes Central vacuole Golgi apparatus Microfilaments Intermediate filaments Microtubules Mitochondrion Peroxisome Chloroplast Plasma membrane Cell wall Plasmodesmata Wall of adjacent cell CYTOSKELETON Structures in the Cell • The tiny organs found inside the cell are called organelles. • Each of these structures performs a specific function that allows the cell to survive. Major Cellular Components • Nucleus • Ribosomes • Endoplasmic reticulum – Smooth and Rough • Golgi body • Various vesicles • Mitochondria • Cytoskeleton Structures found in both Plant and Animal Cells 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. Plasma membrane Nucleus Chromatin Nucleolus Ribosomes Endoplasmic Reticulum Golgi Apparatus Mitochondria Peroxisomes Cytoskeleton Centrosomes Structures Associated with Animal Cells 1. 2. 3. 4. 5. 6. 7. Lysosomes Centrioles Flagella Extracellular matrix Tight Junctions Desmosomes Gap Junctions Structures Associated with Plant Cells 1. 2. 3. 4. Central Vacuoles Chloroplasts Cell Wall Plasmodesmata Components of the Nucleus • Nuclear envelope - Surrounds nucleus • Chromosome - One DNA molecule and associated proteins. Organized DNA. • Chromatin - DNA molecules and histone proteins. Condenses to form DNA. • Nucleolus - RNA and proteins that will be assembled into ribosomal subunits. Cells may have more than one. The Nuclear Envelope • Double - membrane system – Two lipid bilayers. 20-40 nm thick. – Surrounds chromatin/nucleoplasm • Pores allow exchange. Composed of about 100 proteins. Ribosomes • Smallest, most numerous organelle. • Composed of rRNA and proteins. Synthesized by nucleolus. • Large and small subunits. • Found free and bound to E.R. Differ only in what they are making. • Catalyzes formation of peptide bonds. The Endomembrane System • Organelles in which lipids are assembled and proteins are produced and modified • Are in direct contact or send vesicles (membranebound sacs). • Occupy ½ of cell volume. • Nuclear envelope, endoplasmic reticulum, golgi apparatus, lysosomes, vacuole The Endoplasmic Reticulum • Network of tubes and sacs that are continuous with nuclear membrane. Most extensive mem. Sys. • Rough (ribosome studded) and Smooth. – Rough: production of secretory proteins. Signal sequence on polypeptide instructs ribosome to attach to ER. – Smooth: Lipids production, CH2O metabolism, storage of ions, detoxification of drugs/alcohol Fig. 6-11 Cytosol Endoplasmic reticulum (ER) Free ribosomes Bound ribosomes Large subunit 0.5 µm TEM showing ER and ribosomes Small subunit Diagram of a ribosome Fig. 6-10 Nucleus 1 µm Nucleolus Chromatin Nuclear envelope: Inner membrane Outer membrane Nuclear pore Pore complex Surface of nuclear envelope Rough ER Ribosome 1 µm 0.25 µm Close-up of nuclear envelope Pore complexes (TEM) Nuclear lamina (TEM) Fig. 6-12 Smooth ER Rough ER ER lumen Cisternae Ribosomes Transport vesicle Smooth ER Nuclear envelope Transitional ER Rough ER 200 nm Fig. 6-16-1 Nucleus Rough ER Smooth ER Plasma membrane Fig. 6-16-2 Nucleus Rough ER Smooth ER cis Golgi trans Golgi Plasma membrane Fig. 6-16-3 Nucleus Rough ER Smooth ER cis Golgi trans Golgi Plasma membrane Golgi Bodies • Enzymatic finishes on proteins and lipids, and packaging in vesicles. Trans (exit) face • Polarity of cisternae. • Forms glycolipids, glycoproteins, • Products of Golgi leave as vessicles. From one cisternae to another or out of cell. Cis (forming) face Lysosomes • Membrane-bound organelle that contains hydrolytic enzymes responsible for the digestion of macromolecules, autolysis, intracellular digestion. • Dead cells no longer able to maintain H+ gradient (use H+ pump) so organelle breaks down releasing contents. • Made by ER and Golgi. Lysosome Function and Production From Production to Export Vacuoles • Storage of water or ions, pigments, hold food, pump out water. • Are larger than vesicles formed from golgi/E.R. • In plants is enclosed by Tonoplast (membrane) and provides cell with hydrostatic pressure Peroxisomes • Contain enzymes (catalase) that break down H2O2 formed during metabolism of alcohols, F.A.’s. • Specialized forms [glyoxysome] found in seeds and function during germination. Mitochondria • Production of ATP • Double-membrane system – Two distinct compartments • Have their own DNA. Maternal in origin. • Divide on their own, independent of cell. • Have ribosomes, produce enzymes necessary for ATP production. Chloroplast • Found in photosynthetic eukaryotes • Two outer membranes • Semifluid stroma; site of carbon fixation. • Inner thylakoid membrane system; converts l.e. into c.e. • Photosynthetic pigments found in other plastids. Cytoskeleton • Protein fibers that support and give shape to a cell, involved in organelle movement throughout cell, chromosome movement during cell division and large cell movements (cell motility and cytokinesis) • 3 Groups of Fibers classified according to size: – Mircrotubules (thickest) – Intermediated filaments (middle sized) – Microfilaments (thinnest) Components of the Cytoskeleton • Microtubules – and Tubulin subunits; form hollow tube. – Provide framework for cell, organized by centrosome from which they usu. originate. – “Rail” system for organelle transport. • Component of Centriole. – Replicated prior to mitosis. • Form Cilia and Flagella. – 9 + 2 arrangement (eukaryotic characteristic) Fig. 6-UN3 Cilia and Flagella and the Structural Basis of Cell Motility • Surrounded by plasma membrane. • Motor proteins (dynein) on microtubules use ATP to change shape and “ratchet” past one another. • Movement causes bending of cilia/flagella. Components of the Cytoskeleton • Microfilaments (aka actin filaments) – Solid “rope”of two actin proteins – Thinner and more flexible than microtubules – Principle component of muscle fibers. – Provide mechanism to support cell shape. Found just inside the c. mem. – Enable cell movement, phagocytosis and cytokinesis. Components of the Cytoskeleton • Intermediate Filaments – Tough and durable; made of keratin. – Mechanically strengthen/reinforce cells or cell parts that are under stresses. • Provide structure to long cells. • Found in desmosomes. • Give nucleus shape (nuclear lamina) Cell-to Cell Junctions • Plants – Plasmodesmata • Perforations in cell wall that allow passage for water/solutes to adjacent cells. • Animals – Tight Junctions. Prevent leakage between cells (ie. Stomach) – Desmosomes. Mechanically attach cells to each other. Serve as anchoring sites for inter. filaments in cell. – Gap Junctions. Analogous to plasmodesma. Fxn as comm. Pathway between cells. Cardiac muscle, nerves. Intracellular Junctions Fig. 6-32 Tight junction Tight junctions prevent fluid from moving across a layer of cells 0.5 µm Tight junction Intermediate filaments Desmosome Gap junctions Space between cells Plasma membranes of adjacent cells Desmosome 1 µm Extracellular matrix Gap junction 0.1 µm Fig. 6-32a Tight junctions prevent fluid from moving across a layer of cells Tight junction Intermediate filaments Desmosome Gap junctions Space between cells Plasma membranes of adjacent cells Extracellular matrix Fig. 6-32b Tight junction 0.5 µm Fig. 6-32c Desmosome 1 µm Fig. 6-32d Gap junction 0.1 µm Plant Cell Walls • Protect plants, allow for shape and prevent excess H2O uptake. • Composed of cellulose • Plasmodesmata connect neighboring cells. • Secondary cell wall inside of primary wall. Forms wood Plant Cell Wall • Cell secretions form pectin (polysaccharide glue) which acts as adhesive. Laid down in middle lamella to hold cells together. Fig. 6-31 Cell walls Interior of cell Interior of cell 0.5 µm Plasmodesmata Plasma membranes Plasma Membrane • • • • • Forms the boundary for a cell Selectively permeable Made up of phospholipids Fluid mosaic of lipids and proteins Integral proteins are embedded in the membrane • Peripheral proteins are loosely bound to the membrane’s surface • Carbohydrates on the membrane’s surface are crucial in cell to cell recognition • Found in both plant and animal cells Fig. 6-7 Outside of cell Inside of cell 0.1 µm (a) TEM of a plasma membrane Carbohydrate side chain Hydrophilic region Hydrophobic region Hydrophilic region Phospholipid Proteins (b) Structure of the plasma membrane Fig. 6-30 Collagen Proteoglycan complex EXTRACELLULAR FLUID Polysaccharide molecule Carbohydrates Fibronectin Core protein Integrins Proteoglycan molecule Plasma membrane Proteoglycan complex Microfilaments CYTOPLASM Fig. 6-30a Collagen Proteoglycan complex EXTRACELLULAR FLUID Fibronectin Integrins Plasma membrane Microfilaments CYTOPLASM Extracellular matrix (ECM) • Intricate network of proteins and polysaccharides that are organized into a meshwork on the outside of cells. • Large polysaccharides and proteoglygans form a “gellike” material that resist compression. • Proteins like collagen (most abundant protein in animals as part of bone and skin) and elastin (stretch and recoil) provide structure and strength. • Adhesive-like proteins (fibronectins and laminin) help cells attach to the appropriate part of the ECM. Extracellular matrix (ECM) Table 6-1 10 µm 10 µm 10 µm Column of tubulin dimers Keratin proteins Actin subunit Fibrous subunit (keratins coiled together) 25 nm 7 nm Tubulin dimer 8–12 nm Table 6-1a 10 µm Column of tubulin dimers 25 nm Tubulin dimer Table 6-1b 10 µm Actin subunit 7 nm Table 6-1c 5 µm Keratin proteins Fibrous subunit (keratins coiled together) 8–12 nm Fig. 6-UN1 Cell Component Concept 6.3 The eukaryotic cell’s genetic instructions are housed in the nucleus and carried out by the ribosomes Structure Surrounded by nuclear envelope (double membrane) perforated by nuclear pores. The nuclear envelope is continuous with the endoplasmic reticulum (ER). Nucleus Function Houses chromosomes, made of chromatin (DNA, the genetic material, and proteins); contains nucleoli, where ribosomal subunits are made. Pores regulate entry and exit of materials. (ER) Two subunits made of riboProtein synthesis somal RNA and proteins; can be free in cytosol or bound to ER Ribosome Concept 6.4 The endomembrane system regulates protein traffic and performs metabolic functions in the cell Concept 6.5 Mitochondria and chloroplasts change energy from one form to another Extensive network of membrane-bound tubules and sacs; membrane separates lumen from cytosol; continuous with the nuclear envelope. Smooth ER: synthesis of lipids, metabolism of carbohydrates, Ca2+ storage, detoxification of drugs and poisons Golgi apparatus Stacks of flattened membranous sacs; has polarity (cis and trans faces) Modification of proteins, carbohydrates on proteins, and phospholipids; synthesis of many polysaccharides; sorting of Golgi products, which are then released in vesicles. Lysosome Membranous sac of hydrolytic enzymes (in animal cells) Vacuole Large membrane-bounded vesicle in plants Digestion, storage, waste disposal, water balance, cell growth, and protection Mitochondrion Bounded by double membrane; inner membrane has infoldings (cristae) Cellular respiration Endoplasmic reticulum (Nuclear envelope) Chloroplast Peroxisome Rough ER: Aids in synthesis of secretory and other proteins from bound ribosomes; adds carbohydrates to glycoproteins; produces new membrane Breakdown of ingested substances, cell macromolecules, and damaged organelles for recycling Typically two membranes Photosynthesis around fluid stroma, which contains membranous thylakoids stacked into grana (in plants) Specialized metabolic compartment bounded by a single membrane Contains enzymes that transfer hydrogen to water, producing hydrogen peroxide (H2O2) as a by-product, which is converted to water by other enzymes in the peroxisome Fig. 6-UN1a Structure Cell Component Concept 6.3 The eukaryotic cell’s genetic instructions are housed in the nucleus and carried out by the ribosomes Nucleus Function Surrounded by nuclear envelope (double membrane) perforated by nuclear pores. The nuclear envelope is continuous with the endoplasmic reticulum (ER). Houses chromosomes, made of chromatin (DNA, the genetic material, and proteins); contains nucleoli, where ribosomal subunits are made. Pores regulate entry and exit os materials. Two subunits made of ribosomal RNA and proteins; can be free in cytosol or bound to ER Protein synthesis (ER) Ribosome Fig. 6-UN1b Cell Component Concept 6.4 Endoplasmic reticulum The endomembrane system (Nuclear regulates protein traffic and envelope) performs metabolic functions in the cell Golgi apparatus Lysosome Vacuole Structure Function Extensive network of membrane-bound tubules and sacs; membrane separates lumen from cytosol; continuous with the nuclear envelope. Smooth ER: synthesis of lipids, metabolism of carbohydrates, Ca2+ storage, detoxification of drugs and poisons Stacks of flattened membranous sacs; has polarity (cis and trans faces) Rough ER: Aids in sythesis of secretory and other proteins from bound ribosomes; adds carbohydrates to glycoproteins; produces new membrane Modification of proteins, carbohydrates on proteins, and phospholipids; synthesis of many polysaccharides; sorting of Golgi products, which are then released in vesicles. Breakdown of ingested subMembranous sac of hydrolytic stances cell macromolecules, enzymes (in animal cells) and damaged organelles for recycling Large membrane-bounded vesicle in plants Digestion, storage, waste disposal, water balance, cell growth, and protection Fig. 6-UN1c Cell Component Concept 6.5 Mitochondrion Mitochondria and chloroplasts change energy from one form to another Structure Bounded by double membrane; inner membrane has infoldings (cristae) Function Cellular respiration Chloroplast Typically two membranes around fluid stroma, which contains membranous thylakoids stacked into grana (in plants) Photosynthesis Peroxisome Specialized metabolic compartment bounded by a single membrane Contains enzymes that transfer hydrogen to water, producing hydrogen peroxide (H2O2) as a by-product, which is converted to water by other enzymes in the peroxisome