1. Membrane Transport 膜转运 Xia Qiang （夏强）, PhD Department of Physiology Room C518, Block C, Research Building, School of Medicine Tel: 88208252 Email: xiaqiang@zju.edu.cn

2. Objective How molecules and ions get and out of cells?

3. Organelles have their own membranes

4. Cell Membrane (plasma membrane)

5. Phospholipid bilayer ▫Lipids ▫Proteins ▫Carbohydrates

7. Membrane proteins Integral (intrinsic) proteins Peripheral (extrinsic) proteins Integral protein Peripheral protein

8. Functions of membrane proteins Adhesion Some glycoproteins attach to the cytoskeleton and extracellular matrix.

9. Carbohydrates GlycoproteinGlycolipid

10. Membrane Transport Lipid Bilayer -- primary barrier, selectively permeable

11. Membrane Transport ▫Simple Diffusion （单纯扩散） ▫Facilitated Diffusion （易化扩散） ▫Active Transport （主动转运） ▫Endocytosis and Exocytosis （出胞与入胞）

12. Over time, solute molecules placed in a solvent will evenly distribute themselves. START: Initially higher concentration of molecules randomly move toward lower concentration. Diffusional equilibrium is the result (Part b).

13. At time B, some glucose has crossed into side 2 as some cross into side 1.

14. Note: the partition between the two compartments is a membrane that allows this solute to move through it. Net flux accounts for solute movements in both directions.

15. Simple Diffusion Relative to the concentration gradient movement is DOWN the concentration gradient ONLY (higher concentration to lower concentration) Rate of diffusion depends on The concentration gradient Charge on the molecule Size Lipid solubility

16. Facilitated Diffusion ▫Carrier-mediated

17. In simple diffusion, flux rate is limited only by the concentration gradient. In carrier- mediated transport, the number of available carriers places an upper limit on the flux rate.

18. Characteristics of carrier-mediated diffusion net movement always depends on the concentration gradient ▫Specificity ▫Saturation ▫Competition

19. –Channel-mediated 3 cartoon models of integral membrane proteins that function as ion channels; the regulated opening and closing of these channels is the basis of how neurons function.

20. The opening and closing of ion channels results from conformational changes in integral proteins. Discovering the factors that cause these changes is key to understanding excitable cells.

21. Characteristics of ion channels ▫Specificity ▫Gating （门控）

22. Three types of passive, non-coupled transport through integral membrane proteins

25. Outside Inside NH 2 CO 2 H IIIIIIIV ++++++ ++++++ ++++++ ++++++ Voltage-gated Channel –e.g. Voltage-dependent Na + channel

26. Na + channel

27. Balloonfish or fugu

28. Closed Activated Inactivated Na + channel conformation –Open-state –Closed-state

29. Ligand-gated Channel –e.g. N 2 -ACh receptor channel

30. Aquaporin Aquaporins are water channels that exclude ions Aquaporins are found in essentially all organisms, and have major biological and medical importance

31. The Nobel Prize in Chemistry 2003 "for discoveries concerning channels in cell membranes" Peter AgreRoderick MacKinnon "for the discovery of water channels" "for structural and mechanistic studies of ion channels"

32. The dividing wall between the cell and the outside world – including other cells – is far from being an impervious shell. On the contrary, it is perforated by various channels. Many of these are specially adapted to one specific ion or molecule and do not permit any other type to pass. Here to the left we see a water channel and to the right an ion channel.

33. Peter Agre ’ s experiment with cells containing or lacking aquaporin. The aquaporin is necessary for making the cell absorb water and swell

34. Passage of water molecules through the aquaporin AQP1. Because of the positive charge at the center of the channel, positively charged ions such as H3O+, are deflected. This prevents proton leakage through the channel.

35. Model for water permeation through aquaporin

36. In both simple and facilitated diffusion, solutes move in the direction predicted by the concentration gradient. In active transport, solutes move opposite to the direction predicted by the concentration gradient. membrane

37. Active transport Primary active transport （原发性主动转运） Secondary active transport （继发性主动转运）

38. Primary Active Transport making direct use of energy derived from ATP to transport the ions across the cell membrane

39. Here, in the operation of the Na + -K + -ATPase, also known as the “sodium pump,” each ATP hydrolysis moves three sodium ions out of, and two potassium ions into, the cell.

40. Na-K Pump electrogenic pump （生电性泵）

43. Physiological role of Na + -K + pump –Maintaining the Na + and K + gradients across the cell membrane –Partly responsible for establishing a negative electrical potential inside the cell –Controlling cell volume –Providing energy for secondary active transport

44. Other primary active transport Primary active transport of calcium Primary active transport of hydrogen ions

45. Secondary Active Transport The ion gradients established by primary active transport permits the transport of other substances against their concentration gradients

46. Cotransport （同向转运） the ion and the second solute cross the membrane in the same direction (e.g. Na + -glucose, Na + -amino acid cotransport) Countertransport （逆向转运） the ion and the second solute move in opposite directions (e.g. Na + -Ca 2+, Na + -H + exchange)

47. Cotransporters

48. Exchangers

49. Ion gradients, channels, and transporters in a typical cell

52. Here, water is the solvent. The addition of solute lowers the water concentration. Addition of more solute would increase the solute concentration and further reduce the water concentration. Solvent + Solute = Solution Osmosis （渗透）

53. Begin: The partition between the compartments is permeable to water and to the solute. After diffusional equilibrium has occurred: Movement of water and solutes has equalized solute and water concentrations on both sides of the partition.

54. Begin: The partition between the compartments is permeable to water only. After diffusional equilibrium has occurred: Movement of water only has equalized solute concentration.

56. Role of Na-K pump in maintaining cell volume

57. Response to cell shrinking

58. Response to cell swelling

59. Endocytosis and Exocytosis

60. Alternative functions of endocytosis: 1.Transcellular transport 2.Endosomal processing 3.Recycling the membrane 4.Destroying engulfed materials

61. Endocytosis

62. Exocytosis

63. Two pathways of exocytosis Constitutive exocytosis pathway -- Many soluble proteins are continually secreted from the cell by the constitutive secretory pathway Regulated exocytosis pathway -- Selected proteins in the trans Golgi network are diverted into secretory vesicles, where the proteins are concentrated and stored until an extracellular signal stimulates their secretion

64. Steps to exocytosis Vesicle trafficking: In this first step, the vesicle containing the waste product or chemical transmitter is transported through the cytoplasm towards the part of the cell from which it will be eliminated Vesicle tethering: As the vesicle approaches the cell membrane, it is secured and pulled towards the part of the cell from which it will be eliminated Vesicle docking: In this step, the vesicle comes in contact with the cell membrane, where it begins to chemical and physically merge with the proteins in the cell membrane Vesicle priming: In those cells where chemical transmitters are being released, this step involves the chemical preparations for the last step of exocytosis Vesicle fusion: In this last step, the proteins forming the walls of the vesicle merge with the cell membrane and breach, pushing the vesicle contents (waste products or chemical transmitters) out of the cell. This step is the primary mechanism for the increase in size of the cell's plasma membrane

65. Epithelial Transport （上皮转运）

66. Measurement of voltage in an epithelium

67. Models of epithelial solute transport

68. Mechanisms of intestinal glucose absorption

71. Models of “isotonic” water transport in a leaky epithelium

73. Glands

74. Summary Simple Diffusion Facilitated Diffusion Active Transport Endocytosis and Exocytosis Epithelial Transport