Lodish H. - Molecular Cell Biology (5ed, Freeman, 2003) (794361), страница 82
Текст из файла (страница 82)
When viewed by electron microscopy, the lipid bilayer isoften described as looking like a railroad track. Explain howthe structure of the bilayer creates this image.2. Biomembranes contain many different types of lipidmolecules. What are the three main types of lipid moleculesfound in biomembranes? How are the three types similar,and how are they different?use to isolate cells and organelles from complex mixtures,and how do these techniques work?10.
Isolation of some membrane proteins requires the useof detergents; isolation of others can be accomplished withthe use of high-salt solutions. What types of membrane proteins require detergents as part of the isolation procedure?What types of membrane proteins may be isolated with highsalt solutions? Describe how the chemical properties ofdetergents and high salt facilitate the isolation process ofeach type of membrane protein.11. Three systems of cytoskeletal filaments exist in mosteukaryotic cells.
Compare them in terms of composition,function, and structure.12. Individual cytoskeletal filaments are typically organizedinto more complex structures within the cytosol. What twogeneral types of structures do individual filaments combineto form in the cytosol? How are these structures created andmaintained?13. Both light and electron microscopy are commonly usedto visualize cells, cell structures, and the location of specificmolecules. Explain why a scientist may choose one or theother microscopy technique for use in research.3. Lipid bilayers are considered to be two-dimensional fluids; what does this mean? What drives the movement of lipidmolecules and proteins within the bilayer? How can suchmovement be measured? What factors affect the degree ofmembrane fluidity?14.
Why are chemical stains required for visualizing cellsand tissues with the basic light microscope? What advantagedoes fluorescent microscopy provide in comparison to thechemical dyes used to stain specimens for light microscopy?What advantages do confocal scanning microscopy anddeconvolution microscopy provide in comparison to conventional fluorescence microscopy?4. Explain the following statement: The structure of allbiomembranes depends on the chemical properties of phospholipids, whereas the function of each specific biomembrane depends on the specific proteins associated with thatmembrane.15.
In certain electron microscopy methods, the specimenis not directly imaged. How do these methods provide information about cellular structure, and what types of structuresdo they visualize?5. Name the three groups into which membrane-associatedproteins may be classified. Explain the mechanism by whicheach group associates with a biomembrane.6. Although both faces of a biomembrane are composed ofthe same general types of macromolecules, principally lipidsand proteins, the two faces of the bilayer are not identical.What accounts for the asymmetry between the two faces?7. One of the defining features of eukaryotic cells is thepresence of organelles.
What are the major organelles of eukaryotic cells, and what is the function of each? What is thecytosol? What cellular processes occur within the cytosol?8. Cell organelles such as mitochondria, chloroplasts, andthe Golgi apparatus each have unique structures. How is thestructure of each organelle related to its function?9. Much of what we know about cellular function dependson experiments utilizing specific cells and specific parts (e.g.,organelles) of cells. What techniques do scientists commonlyA N A LY Z E T H E DATAMouse liver cells were homogenized and the homogenatesubjected to equilibrium density-gradient centrifugation withsucrose gradients. Fractions obtained from these gradientswere assayed for marker molecules (i.e., molecules that arelimited to specific organelles).
The results of these assays areshown in the figure. The marker molecules have the following functions: Cytochrome oxidase is an enzyme involved inthe process by which ATP is formed in the complete aerobicdegradation of glucose or fatty acids; ribosomal RNA formspart of the protein-synthesizing ribosomes; catalase catalyzesdecomposition of hydrogen peroxide; acid phosphatasehydrolyzes monophosphoric esters at acid pH; cytidylyltransferase is involved in phospholipid biosynthesis; andamino acid permease aids in transport of amino acids acrossmembranes.ReferencesA100BCD ESprong, H., P. van der Sluijs, and G.
van Meer. 2001. How proteins move lipids and lipids move proteins. Nature Rev. Mol. CellBiol. 2:504–513.Tamm, L. K., V. K. Kiessling, and M. L. Wagner. 2001. Membrane dynamics. Encyclopedia of Life Sciences. Nature PublishingGroup.Vance, D. E., and J. E. Vance. 2002. Biochemistry of Lipids,Lipoproteins, and Membranes, 4th ed. Elsevier.Yeager, P.
L. 2001. Lipids. Encyclopedia of Life Sciences. Nature Publishing Group.F% of maximum8060Biomembranes: Protein Componentsand Basic Functions402001950510Fraction number50%Curve A = cytochrome oxidaseCurve B = ribosomal RNACurve C = catalase1520Sucrose 0%Curve D = acid phosphataseCurve E = cytidylyl transferaseCurve F = amino acid permeasea.
Name the marker molecule and give the number ofthe fraction that is most enriched for each of the following: lysosomes; peroxisomes; mitochondria; plasma membrane; rough endoplasmic reticulum; smooth endoplasmicreticulum.b. Is the rough endoplasmic reticulum more or less densethan the smooth endoplasmic reticulum? Why?c. Describe an alternative approach by which you couldidentify which fraction was enriched for which organelle.d.
How would addition of a detergent to the homogenateaffect the equilibrium density-gradient results?REFERENCESGeneral Histology Texts and AtlasesCross, P. A., and K. L. Mercer. 1993. Cell and Tissue Ultrastructure: A Functional Perspective. W.
H. Freeman and Company.Fawcett, D. W. 1993. Bloom and Fawcett: A Textbook of Histology, 12th ed. Chapman & Hall.Kessel, R., and R. Kardon. 1979. Tissues and Organs: A TextAtlas of Scanning Electron Microscopy. W. H. Freeeman andCompany.Biomembranes: Lipid Compositionand Structural OrganizationSimons, K., and D. Toomre. 2000. Lipid rafts and signal transduction. Nature Rev. Mol. Cell Biol.
1:31–41.Cullen, P. J., G. E. Cozier, G. Banting, and H. Mellor. 2001.Modular phosphoinositide-binding domains: their role in signallingand membrane trafficking. Curr. Biol. 11:R882–R893.Lanyi, J. K., and H. Luecke. 2001. Bacteriorhodopsin. Curr.Opin.
Struc. Biol. 11:415–519.MacKenzie, K. R., J. H. Prestegard, and D. M. Engelman. 1997.A transmembrane helix dimer: structure and implications. Science276:131–133.Minor, D. L. 2001. Potassium channels: life in the post-structuralworld. Curr. Opin. Struc. Biol. 11:408–414.Schulz, G. E. 2000. -Barrel membrane proteins. Curr.
Opin.Struc. Biol. 10:443–447.Organelles of the Eukaryotic CellBainton, D. 1981. The discovery of lysosomes. J. Cell Biol.91:66s–76s.Cuervo, A. M., and J. F. Dice. 1998. Lysosomes: a meeting pointof proteins, chaperones, and proteases. J. Mol. Med. 76:6–12.de Duve, C. 1996. The peroxisome in retrospect. Ann. NY Acad.Sci. 804:1–10.Holtzman, E. 1989. Lysosomes.
Plenum Press.Lamond, A., and W. Earnshaw. 1998. Structure and function inthe nucleus. Science 280:547–553.Masters, C., and D. Crane. 1996. Recent developments in peroxisome biology. Endeavour 20:68–73.Palade, G. 1975. Intracellular aspects of the process of proteinsynthesis. Science 189:347–358. The Nobel Prize lecture of a pioneerin the study of cellular organelles. (See also de Duve, 1996.)Subramani, S. 1998. Components involved in peroxisome import, biogenesis, proliferation, turnover, and movement. Physiol.
Rev.78:171–188.The Cytoskeleton: Components and Structural FunctionsBray, D. 2001. Cell Movements: From Molecules to Motility.Garland. Excellent overview of the cytoskeleton and motility.Various authors. Curr. Topics Cell Biol. February issue is alwaysdevoted to the cytoskeleton.Purification of Cells and Their PartsBattye, F. L., and K. Shortman.
1991. Flow cytometry and cellseparation procedures. Curr. Opin. Immunol. 3:238–241.de Duve, C. 1975. Exploring cells with a centrifuge. Science189:186–194. The Nobel Prize lecture of a pioneer in the study ofcellular organelles.de Duve, C., and H. Beaufay. 1981. A short history of tissuefractionation. J. Cell Biol. 91:293s–299s.Howell, K. E., E.
Devaney, and J. Gruenberg. 1989. Subcellularfractionation of tissue culture cells. Trends Biochem. Sci. 14:44–48.196CHAPTER 5 • Biomembranes and Cell ArchitectureOrmerod, M. G., ed. 1990. Flow Cytometry: A Practical Approach. IRL Press.Rickwood, D. 1992. Preparative Centrifugation: A Practical Approach. IRL Press.Visualizing Cell ArchitectureBastiaens, P. I.