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Lecture 14 plasma membrane transport Active transport pp73-77

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1 Lecture 14 plasma membrane transport Active transport pp73-77

2 Active Processes Whenever a cell uses the bond energy of ATP to move solutes across the membrane, the process is referred to as active. Substances moved actively across the plasma membrane are usually unable to pass in the necessary direction by passive transport processes. The substance may be too large to pass through the channels, incapable of dissolving in the lipid bilayer, or unable to move down its concentration gradient.

3 Membrane Transport: Active Processes
Two types of active processes: Active transport Vesicular transport Both use ATP to move solutes across a living plasma membrane

4 Active transport    like carrier-mediated facilitated diffusion, requires carrier proteins that combine specifically and reversibly with the transported substances. However, facilitated diffusion always honors concentration gradients because its driving force is kinetic energy. In contrast, the active transporters or solute pumps move solutes, most importantly ions (such as Na+, K+, and Ca2+), “uphill” against a concentration gradient. To do this work, cells must expend the energy ofATP.

5 (b) Carrier-mediated facilitated diffusion via a protein
Lipid-insoluble solutes (such as sugars or amino acids) (b) Carrier-mediated facilitated diffusion via a protein carrier specific for one chemical; binding of substrate causes shape change in transport protein Figure 3.7b

6 Active Transport Requires carrier proteins (solute pumps)
Moves solutes against a concentration gradient Types of active transport: Primary active transport Secondary active transport

7 Primary Active Transport
Energy from hydrolysis of ATP causes shape change in transport protein so that bound solutes (ions) are “pumped” across the membrane

8 Primary Active Transport
Sodium-potassium pump (Na+-K+ ATPase) Located in all plasma membranes Involved in primary and secondary active transport of nutrients and ions Maintains electrochemical gradients essential for functions of muscle and nerve tissues

9 Figure 3.10 Extracellular fluid Na+ Na+-K+ pump ATP-binding site K+
Na+ bound Cytoplasm 1 Cytoplasmic Na+ binds to pump protein. P ATP K+ released ADP 6 K+ is released from the pump protein and Na+ sites are ready to bind Na+ again. The cycle repeats. 2 Binding of Na+ promotes phosphorylation of the protein by ATP. Na+ released K+ bound P Pi K+ 5 K+ binding triggers release of the phosphate. Pump protein returns to its original conformation. 3 Phosphorylation causes the protein to change shape, expelling Na+ to the outside. P 4 Extracellular K+ binds to pump protein. Figure 3.10

10 1 Extracellular fluid Na+ Na+-K+ pump K+ ATP-binding site Cytoplasm
Cytoplasmic Na+ binds to pump protein. Figure 3.10 step 1

11 Na+ bound P ATP ADP 2 Binding of Na+ promotes phosphorylation of the protein by ATP. Figure 3.10 step 2

12 Na+ released P Phosphorylation causes the protein to change shape, expelling Na+ to the outside. 3 Figure 3.10 step 3

13 K+ P 4 Extracellular K+ binds to pump protein. Figure 3.10 step 4

14 K+ bound Pi K+ binding triggers release of the phosphate. Pump protein returns to its original conformation. 5 Figure 3.10 step 5

15 K+ released 6 K+ is released from the pump protein and Na+ sites are ready to bind Na+ again. The cycle repeats. Figure 3.10 step 6

16 Figure 3.10 Extracellular fluid Na+ Na+-K+ pump ATP-binding site K+
Na+ bound Cytoplasm 1 Cytoplasmic Na+ binds to pump protein. P ATP K+ released ADP 6 K+ is released from the pump protein and Na+ sites are ready to bind Na+ again. The cycle repeats. 2 Binding of Na+ promotes phosphorylation of the protein by ATP. Na+ released K+ bound P Pi K+ 5 K+ binding triggers release of the phosphate. Pump protein returns to its original conformation. 3 Phosphorylation causes the protein to change shape, expelling Na+ to the outside. P 4 Extracellular K+ binds to pump protein. Figure 3.10

17 Secondary Active Transport
Depends on an ion gradient created by primary active transport Energy stored in ionic gradients is used indirectly to drive transport of other solutes

18 Secondary Active Transport
Cotransport—always transports more than one substance at a time Symport system: Two substances transported in same direction Antiport system: Two substances transported in opposite directions

19 Symport carries 2 or more solutes thru the membrane simultaneously in the SAME direction Cotransport = process

20 Antiport Carriers 2 or more solutes in the OPPOSITE directions
Counter transport = process

21 The ATP-driven Na+-K+ pump stores energy by creating a
Extracellular fluid Glucose Na+-glucose symport transporter loading glucose from ECF Na+-glucose symport transporter releasing glucose into the cytoplasm Na+-K+ pump Cytoplasm 1 The ATP-driven Na+-K+ pump stores energy by creating a steep concentration gradient for Na+ entry into the cell. 2 As Na+ diffuses back across the membrane through a membrane cotransporter protein, it drives glucose against its concentration gradient into the cell. (ECF = extracellular fluid) Figure 3.11

22 The ATP-driven Na+-K+ pump stores energy by creating a
Extracellular fluid Na+-K+ pump Cytoplasm 1 The ATP-driven Na+-K+ pump stores energy by creating a steep concentration gradient for Na+ entry into the cell. Figure 3.11 step 1

23 The ATP-driven Na+-K+ pump stores energy by creating a
Extracellular fluid Glucose Na+-glucose symport transporter loading glucose from ECF Na+-glucose symport transporter releasing glucose into the cytoplasm Na+-K+ pump Cytoplasm 1 The ATP-driven Na+-K+ pump stores energy by creating a steep concentration gradient for Na+ entry into the cell. 2 As Na+ diffuses back across the membrane through a membrane cotransporter protein, it drives glucose against its concentration gradient into the cell. (ECF = extracellular fluid) Figure 3.11 step 2

24 Vesicular Transport Transport of large particles, macromolecules, and fluids across plasma membranes Requires cellular energy (e.g., ATP)

25 Vesicular Transport Functions: Exocytosis—transport out of cell
Endocytosis—transport into cell Transcytosis—transport into, across, and then out of cell Substance (vesicular) trafficking—transport from one area or organelle in cell to another

26 Endocytosis and Transcytosis
Involve formation of protein-coated vesicles Often receptor mediated, therefore very selective

27 Types Endocytosis – vesicular process that brings matter INTO the cell
Exocytosis – vesicular process that releases matter OUTSIDE the cell

28 Exocytosis Figure 3.12a

29

30 Vesicular Transport Transcytosis – moving substances into, across, and then out of a cell Vesicular trafficking – moving substances from one area in the cell to another Phagocytosis – pseudopods engulf solids and bring them into the cell’s interior

31 Vesicular Transport Fluid-phase endocytosis – the plasma membrane infolds, bringing extracellular fluid and solutes into the interior of the cell Receptor-mediated endocytosis – clathrin-coated pits provide the main route for endocytosis and transcytosis Non-clathrin-coated vesicles – caveolae that are platforms for a variety of signaling molecules

32 Coated pit ingests substance. Extracellular fluid Plasma membrane
1 Extracellular fluid Plasma membrane Protein coat (typically clathrin) Cytoplasm 2 Protein- coated vesicle detaches. Coat proteins detach and are recycled to plasma membrane. 3 Transport vesicle Endosome Uncoated endocytic vesicle 4 Uncoated vesicle fuses with a sorting vesicle called an endosome. Transport vesicle containing membrane components moves to the plasma membrane for recycling. 5 Lysosome 6 Fused vesicle may (a) fuse with lysosome for digestion of its contents, or (b) deliver its contents to the plasma membrane on the opposite side of the cell (transcytosis). (b) (a) Figure 3.12

33 Coated pit ingests substance. Extracellular fluid Plasma membrane
1 Extracellular fluid Plasma membrane Protein coat (typically clathrin) Cytoplasm Figure 3.12 step 1

34 Coated pit ingests substance. Extracellular fluid Plasma membrane
1 Extracellular fluid Plasma membrane Protein coat (typically clathrin) Cytoplasm 2 Protein- coated vesicle detaches. Figure 3.12 step 2

35 Coated pit ingests substance. Extracellular fluid Plasma membrane
1 Extracellular fluid Plasma membrane Protein coat (typically clathrin) Cytoplasm 2 Protein- coated vesicle detaches. Coat proteins detach and are recycled to plasma membrane. 3 Figure 3.12 step 3

36 Coated pit ingests substance. Extracellular fluid Plasma membrane
1 Coated pit ingests substance. Extracellular fluid Plasma membrane Protein coat (typically clathrin) Cytoplasm 2 Protein- coated vesicle detaches. 3 Coat proteins detach and are recycled to plasma membrane. Endosome Uncoated endocytic vesicle Uncoated vesicle fuses with a sorting vesicle called an endosome. 4 Figure 3.12 step 4

37 Coated pit ingests substance. Extracellular fluid Plasma membrane
1 Coated pit ingests substance. Extracellular fluid Plasma membrane Protein coat (typically clathrin) Cytoplasm 2 Protein- coated vesicle detaches. Coat proteins detach and are recycled to plasma membrane. 3 Transport vesicle Endosome Uncoated endocytic vesicle 4 Uncoated vesicle fuses with a sorting vesicle called an endosome. Transport vesicle containing membrane components moves to the plasma membrane for recycling. 5 Figure 3.12 step 5

38 Coated pit ingests substance. Extracellular fluid Plasma membrane
1 Extracellular fluid Plasma membrane Protein coat (typically clathrin) Cytoplasm 2 Protein- coated vesicle detaches. 3 Coat proteins detach and are recycled to plasma membrane. Transport vesicle Endosome Uncoated endocytic vesicle Uncoated vesicle fuses with a sorting vesicle called an endosome. 4 Transport vesicle containing membrane components moves to the plasma membrane for recycling. 5 Lysosome 6 Fused vesicle may (a) fuse with lysosome for digestion of its contents, or (b) deliver its contents to the plasma membrane on the opposite side of the cell (transcytosis). (b) (a) Figure 3.12 step 6

39 Endocytosis Phagocytosis—pseudopods engulf solids and bring them into cell’s interior Macrophages and some white blood cells

40 The cell engulfs a large particle by forming pro-
Phagocytosis The cell engulfs a large particle by forming pro- jecting pseudopods (“false feet”) around it and en- closing it within a membrane sac called a phagosome. The phagosome is combined with a lysosome. Undigested contents remain in the vesicle (now called a residual body) or are ejected by exocytosis. Vesicle may or may not be protein- coated but has receptors capable of binding to microorganisms or solid particles. Phagosome Figure 3.13a

41 Endocytosis Fluid-phase endocytosis (pinocytosis)—plasma membrane infolds, bringing extracellular fluid and solutes into interior of the cell Nutrient absorption in the small intestine

42 The cell “gulps” drops of extracellular fluid containing
(b) Pinocytosis The cell “gulps” drops of extracellular fluid containing solutes into tiny vesicles. No receptors are used, so the process is nonspecific. Most vesicles are protein-coated. Vesicle Figure 3.13b

43 Endocytosis Receptor-mediated endocytosis—clathrin-coated pits provide main route for endocytosis and transcytosis Uptake of enzymes low-density lipoproteins, iron, and insulin

44 Extracellular substances bind to specific receptor
Receptor-mediated endocytosis Extracellular substances bind to specific receptor proteins in regions of coated pits, enabling the cell to ingest and concentrate specific substances (ligands) in protein-coated vesicles. Ligands may simply be released inside the cell, or combined with a lysosome to digest contents. Receptors are recycled to the plasma membrane in vesicles. Vesicle Receptor recycled to plasma membrane Figure 3.13c

45 Exocytosis Examples: Hormone secretion Neurotransmitter release
Mucus secretion Ejection of wastes

46 Summary of Active Processes
Energy Source Example Primary active transport ATP Pumping of ions across membranes Secondary active transport Ion gradient Movement of polar or charged solutes across membranes Exocytosis Secretion of hormones and neurotransmitters Phagocytosis White blood cell phagocytosis Pinocytosis Absorption by intestinal cells Receptor-mediated endocytosis Hormone and cholesterol uptake Also see Table 3.2

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48

49 Thank you

50 Membrane Potential Separation of oppositely charged particles (ions) across a membrane creates a membrane potential (potential energy measured as voltage) Resting membrane potential (RMP): Voltage measured in resting state in all cells Ranges from –50 to –100 mV in different cells Results from diffusion and active transport of ions (mainly K+)

51 Generation and Maintenance of RMP
The Na+ -K+ pump continuously ejects Na+ from cell and carries K+ back in Some K+ continually diffuses down its concentration gradient out of cell through K+ leakage channels Membrane interior becomes negative (relative to exterior) because of large anions trapped inside cell

52 Generation and Maintenance of RMP
Electrochemical gradient begins to attract K+ back into cell RMP is established at the point where the electrical gradient balances the K+ concentration gradient A steady state is maintained because the rate of active transport is equal to and depends on the rate of Na+ diffusion into cell

53 1 2 3 K+ diffuse down their steep Extracellular fluid
concentration gradient (out of the cell) via leakage channels. Loss of K+ results in a negative charge on the inner plasma membrane face. 1 Extracellular fluid K+ also move into the cell because they are attracted to the negative charge established on the inner plasma membrane face. 2 A negative membrane potential (–90 mV) is established when the movement of K+ out of the cell equals K+ movement into the cell. At this point, the concentration gradient promoting K+ exit exactly opposes the electrical gradient for K+ entry. 3 Potassium leakage channels Protein anion (unable to follow K+ through the membrane) Cytoplasm Figure 3.15

54 Cell-Environment Interactions
Involves glycoproteins and proteins of glycocalyx Cell adhesion molecules (CAMs) Membrane receptors

55 Roles of Cell Adhesion Molecules
Anchor cells to extracellular matrix or to each other Assist in movement of cells past one another CAMs of blood vessel lining attract white blood cells to injured or infected areas Stimulate synthesis or degradation of adhesive membrane junctions Transmit intracellular signals to direct cell migration, proliferation, and specialization

56 Roles of Membrane Receptors
Contact signaling—touching and recognition of cells; e.g., in normal development and immunity Chemical signaling—interaction between receptors and ligands (neurotransmitters, hormones and paracrines) to alter activity of cell proteins (e.g., enzymes or chemically gated ion channels) G protein–linked receptors—ligand binding activates a G protein, affecting an ion channel or enzyme or causing the release of an internal second messenger, such as cyclic AMP

57 Cascade of cellular responses (metabolic and structural changes)
1 Ligand (1st messenger) binds to the receptor. 2 The activated receptor binds to a G protein and activates it. 3 Activated G protein activates (or inactivates) effector protein (e.g., an enzyme) by causing its shape to change. Extracellular fluid Effector protein (e.g., an enzyme) Ligand Receptor 4 Activated effector enzymes catalyze reactions that produce 2nd messengers in the cell Inactive 2nd messenger G protein GDP Active 2nd messenger 5 Second messengers activate other enzymes or ion channels 6 Activated kinase enzymes Kinase enzymes transfer phosphate groups from ATP to specific proteins and activate a series of other enzymes that trigger various cell responses. Cascade of cellular responses (metabolic and structural changes) Intracellular fluid Figure 3.16

58 Figure 3.16 step 1 1 Ligand (1st messenger) binds to the receptor.
Extracellular fluid Ligand Receptor Intracellular fluid Figure 3.16 step 1

59 Figure 3.16 step 2 1 Ligand (1st messenger) binds to the receptor. 2
The activated receptor binds to a G protein and activates it. Extracellular fluid Ligand Receptor G protein GDP Intracellular fluid Figure 3.16 step 2

60 Figure 3.16 step 3 1 Ligand (1st messenger) binds to the receptor. 2
The activated receptor binds to a G protein and activates it. 3 Activated G protein activates (or inactivates) effector protein (e.g., an enzyme) by causing its shape to change. Extracellular fluid Effector protein (e.g., an enzyme) Ligand Receptor G protein GDP Intracellular fluid Figure 3.16 step 3

61 Figure 3.16 step 4 1 Ligand (1st messenger) binds to the receptor. 2
The activated receptor binds to a G protein and activates it. 3 Activated G protein activates (or inactivates) effector protein (e.g., an enzyme) by causing its shape to change. Extracellular fluid Effector protein (e.g., an enzyme) Ligand Receptor 4 Activated effector enzymes catalyze reactions that produce 2nd messengers in the cell Inactive 2nd messenger G protein GDP Active 2nd messenger Intracellular fluid Figure 3.16 step 4

62 Figure 3.16 step 5 1 Ligand (1st messenger) binds to the receptor. 2
The activated receptor binds to a G protein and activates it. 3 Activated G protein activates (or inactivates) effector protein (e.g., an enzyme) by causing its shape to change. Extracellular fluid Effector protein (e.g., an enzyme) Ligand Receptor 4 Activated effector enzymes catalyze reactions that produce 2nd messengers in the cell Inactive 2nd messenger G protein 5 GDP Active 2nd messenger Second messengers activate other enzymes or ion channels Activated kinase enzymes Intracellular fluid Figure 3.16 step 5

63 Cascade of cellular responses (metabolic and structural changes)
1 Ligand (1st messenger) binds to the receptor. 2 The activated receptor binds to a G protein and activates it. 3 Activated G protein activates (or inactivates) effector protein (e.g., an enzyme) by causing its shape to change. Extracellular fluid Effector protein (e.g., an enzyme) Ligand Receptor 4 Activated effector enzymes catalyze reactions that produce 2nd messengers in the cell Inactive 2nd messenger G protein GDP Active 2nd messenger 5 Second messengers activate other enzymes or ion channels 6 Activated kinase enzymes Kinase enzymes transfer phosphate groups from ATP to specific proteins and activate a series of other enzymes that trigger various cell responses. Cascade of cellular responses (metabolic and structural changes) Intracellular fluid Figure 3.16 step 6


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