1. revised 10/24/2013 Biochemistry I Dr. Loren Williams Chapter 10 Membrane Transport

2. Thermodynamics of Transport

9. Mammalian Cell inside 12 mM Na+ 140 mM K+ outside 150 mM Na+ 4mM K+

10. Passive Transport Movement of molecules across membranes driven by electochemicalpotential Mechanism is either simple diffusion or facilitated (mediated) diffusion Not thermodynamically linked to other processes No ATP hydrolysis Mediation of transport across membranes required for charged, polar or large molecules 1) Ionophores lipid-soluble molecules synthesized by microorganisms facilitate transport of ions across membrane. Valinomycin a potassium-specific passive transporter a dodecadepsipeptide antibiotic produced by several Streptomyces strains highly selective for potassium ions over sodium and other ions Kd for potassium is 106 Kd for sodium is 10. facilitates transport of K+ down the electrochemical potential gradient. Deadly

11. Figure 10-1

14. Sodium (Na+) Oxidation state: +1 Ionic radius: 0.95 Å Preferred ligands: O Number of ligands: 6-7 Preferred geometry: octahedral (not a strong preference) Potassium (K+) Oxidation state: +1 Ionic radius: 1.33 Å Preferred ligands: O Number of ligands: 4-7 Preferred geometry: variable, octahedral Calcium (Ca2+) Oxidation state: +2 Ionic radius: 0.99 Å Preferred ligands: O Number of ligands: 6-10 Preferred geometry: variable

15. Passive Transport 2) Ion Channels integral membrane proteins found in the membranes of all cells (necessary to keep cells from exploding) highly selective for specific ions (K+vs Na+vs Ca2+etc) fast: rate of transport is close to the diffusion limit very tightly regulated gated: flow of ions across the cell membrane is turned off or no in response to stimuli Sensors: pH, ligands, voltage, etc K+ channels and anion channels hyperpolarize cells (cause the membrane potential to become more negative), Na+and Ca2+ channels and non-selective cation channels depolarize cells (cause the membrane potential to become more positive). Ion channels form pores that permit the flux of ions down their electrochemical gradient.

16. Mammalian Cell outside (high Na+) 150 mM Na+ 4 mM K+ inside (high K+) 12 mM Na+ 140 mM K+ Ion channels form pores that permit the flux of ions down their electrochemical gradient.

17. KcsAK+ channel

18. KcsA K+ Ion Channel 10,000-fold selectivity of K+ over Na+ homo tetramer four identical protein subunits two transmembrane helices, central pore three parts: a selectivity filter (extracellular side), a dilated water-filled cavity (center), gate (cytoplasmic side, proton-activated, opens at acidic pH)

19. How does ion selectivity work? How is K+ distinguished from Na+? Ion Dehydration: disrupts favorable molecular interactions (DH>0) Ion Coordination: forms favorable molecular interactions (DH<0) To enter the selectivity channel the ions must dehydrate. The enthalpy of ion coordination by the selectivity channel has to offset the unfavorable dehydration enthalpy. Na+is smaller (ionic radius 0.95 Å) than K+(ionic radius 1.33 Å). The coordination geometry in the selectivity channel is bad for Na+: The O-Na+distances are too long. The O-K+distances are just right. Optimum O - Na+distance = 2.4 Å (0.95 + 1.5) Optimum O - K+distance = 2.8 Å (1.33 + 1.5) K+ is about the same size as water, Na+ is smaller than water. Free energy landscape for K+ is featureless throughout the channel.

20. Channels are gated Mechanics (touch, sound, etc) Ligands (neurotransmitters…) Signals (Ca2+…) Voltage (changes in membrane potential)

21. Voltage Gating (Kv Channel): Gate # 1 S4 helix (+++++ charged) Depolarization moves S4 toward the outside (extracelluar side) of the membrane, and opens the channel.

22. Voltage Gating (Kv Channel): Gate # 2 inactivation ball Closes the channel a few msec after the S4 helix opens it. The channel does not reopen until the potential is reset,

23. Outside open closed Inside Figure 10-9

24. Figure 10-9a

25. Figure 10-9b,c

26. Action Potential stimulation of a neuron open Na+ channel depolarize close Na+ channel open K+ channel hyperpolarize close Na+ channel relax

28. fall 2013, not covered Figure 10-3

29. fall 2013, not covered Figure 10-6b

30. fall 2013, not covered Figure 10-10

31. fall 2013, not covered Figure 10-11a

32. fall 2013, not covered Figure 10-11b

33. fall 2013, not covered Figure 10-12

34. Glucose Transporter Basis of selectivity Figure 10-13

35. fall 2013, not covered Box 10-1a

36. fall 2013, not covered Box 10-1b

37. fall 2013, not covered Box 10-2

38. fall 2013, not covered Figure 10-14

39. fall 2013, not covered Page 310

40. fall 2013, not covered Figure 10-15

41. Figure 10-16

42. fall 2013, not covered Page 312

43. The Na+/K+ pump (ATPase) maintains resting potential regulates cell volume, signal transducer/integrator 20% of cell's energy expenditure. 50-70% cell's energy expenditure for neurons Pumps 3 Na+ out for every 2 K+ in (hydrolyzes 1 ATP in the process).

44. unphosphorylated pump binds 3 intracellular Na+ and an ATP, phosphate is transfered from ATP to aspartate of the pump, release of ADP conformational change exposes the Na+ ions to the outside, all Na+ are released outside pump binds 2 extracellular K+ ions. dephosphorylationof the pump, both K+ are released on the inside cycle completed start

45. Box 10-3

46. Figure 10-18

47. Figure 10-19

48. Figure 10-20

49. Figure 10-21

50. Figure 10-21a