Biological Membranes : Problems and solutions 1. Synthesis y of Gastric Juice: Energetics g Gastric juice (pH 1.

5) is produced by pumping HCl from blood plasma (pH 7.4) into the stomach. stomach Calculate the amount of free energy required to concentrate the H+ in 1 L of gastric juice at 37 C. Under cellular ll l conditions, diti h how many moles l of f ATP must be hydrolyzed to provide this amount of free energy? gy The free-energy gy change for ATP hydrolysis under cellular conditions is about - 58 kJ/mol. Ignore the effects of the transmembrane electrical potential. (Problem # 7, page # 415-416, Michael M. C Cox and d David D id L. L Nelson: N l P i i l Principles of f Biochemistry, Fifth Edition)

Solution p pH = log [H] [H], [H] = 10-pH. At pH 1.5, [H] = 10 - 1.5 = 3.2 x 10 - 2 M. At pH 7.4, [H] = 10-7.4 = 4.0 x 10-8 M.

Gt = RT ln (C2/C1)

at 37 C, RT = 2.58 kJ/mol,

Gt = (2.58 kJ/mol) ln (3.2 x 10 = 35 kJ/mol.

-2

/ 4.0 x 10-8 )

The amount of ATP required to provide 35 kJ is 0.60 0 60 mol (35 kJ/58kJ)

2 Energetics of the Na+K+ ATPase 2. For a typical vertebrate cell with a membrane potential of 0.070 V (inside negative), ti ) what h t is i the th free-energy f change h for transporting 1 mol of Na+ from the cell into the blood at 37oC? Assume the concentration of Na+ inside the cell is 12 mM, and that in blood plasma is 145 mM. (Problem # 8, 8 page # 416 416, Michael M. M Cox and David L. Nelson: Principles of Biochemistry, Fifth Edition)

S l ti Solution:

Gt = RT ln (C2/C1) + Z
Z = charge on the ion = Faraday constant (96,480 J/v.mol) =Transmembrane electrical potential (in volts)
Gt = (2.58 kJ/mol) ln (145/12) + (1)(96,480 J/V mol)(0.070 V)

= 6.4 kJ/mol + 6.8 kJ/mol = 13 kJ/mol

6.8 kJ/mol is the membrane potential portion

3. Water Flow through g an Aquaporin q p A human erythrocyte has about 2 x 105, AQP-1 monomers. If water molecules flow through the plasma membrane at a rate of 5x108 per AQP-1 tetramer per second, and the volume of an erythrocyte is 5x10-11 mL, h how rapidly idl could ld an erythrocyte th t halve h l it its volume as it encountered the high osmolarity y( (1 M) ) in the interstitial fluid of the renal medulla? Assume that the erythrocyte consists entirely of water. (Problem # 16, 16 page # 416, 416 Michael M. M Cox and David L. Nelson: Principles of Biochemistry, Fifth Edition)

Solution 1. Calculate the number of water molecules that must leave the erythrocyte to halve its volume. The volume of the cell is 5 x 10-11 ml. For [H2O] = 55 M The number of water molecules in the cell = (5 x 10-11 ml/cell )(6.02 x 1020 molecules/mmol) (55 mmol H2O/ml) =1.7 x 1012 Half of these molecules (8.5 x 1011) must l leave to t h halve l th the cell ll volume. l

2. Calculate how fast the cell can lose water t molecules. l l The cell has 2 x 105 aquaporin monomers, , or 5 x 104 tetramers. Each tetramer allows passage of 5 x 108 H2O molecules per second, The flux of water molecules through the plasma membrane = (5 x 108 H2O molecules/s/aquaporin t t tetramer) ) (5 x 104 aquaporin i tetramers/cell) = 2.5 x 1013 H2O molecules/s Time required to remove half the volume of water = (8.5 x 1011 H2O molecules)/(2.5 x 1013 H2O molecules/s) = 3 x 10-2 s

4. Surface Density of a Membrane Protein E. coli can be induced to make about 10,000 copies of the lactose transporter (Mr 31 000) per cell. 31,000) cell Assume that E. E coli is a cylinder 1 m in diameter and 2 m long. What fraction of the plasma membrane surface is occupied by the lactose transporter molecules? Explain how you arrived a ed at t this s co conclusion. c us o (Problem # 21, page # 417, Michael M. Cox and David L. Nelson: Principles of Biochemistry Fifth Edition) Biochemistry,

Solution Calculate E. coli surface area. The surface area of a cylinder = where , r= radius, d = diameter, and h =h height. i ht For a cylinder 2 m high and 1 m in diameter, the surface area = 2 (0.5 m)2 + (1 m) (2 m) = 8 m2. To estimate the cross cross-sectional sectional area of a globular protein of Mr 31,000, Use the dimensions for hemoglobin (Mr 64,500; diameter 5.5 5 5 nm), thus th s a protein of Mr 31,000 has a diameter of about 3 nm, assuming the proteins have the same density.

The crosssectional area of a sphere p of diameter 3 nm (0.003 m)or of a single transporter molecule = r2 = 3.14(1.5 3 14(1 5 x 10-3 m)2 = 7x10-6 m2 The total cross-sectional area of 10,000 2 m2. t transporter t molecules l l =7 7x10 10-2 The fraction of an E. coli cell surface covered by y transporter p molecules= (7x10-2 m2)/(8 m2) = 0.009, or about 1%.

5. Molecular Species in the E. coli Membrane The plasma membrane of E. coli is about 75% protein and 25% phospholipid by weight. g How many y molecules of membrane lipid are present for each molecule of membrane protein? Assume an average protein Mr of 50,000 50 000 and an average phospholipid Mr of 750. What more would you need to know to estimate the fraction of the membrane surface that is covered by lipids? (Problem # 23, ( , p page g # 417, , Michael M. Cox and David L. Nelson: Principles of Biochemistry, Fifth Edition)

Solution Consider a sample that contains 1 g of membrane, of which 0.75 g is protein (Mr 50,000) and 0.25 g is phospholipid (Mr 750). 5 mol (0.75 g protein)(1 mol/5 x104 g) = 1.5 x10-5 protein in 1 g of membrane (0.25 g p ( phospholipid)(1 p p )( mol/750 g) = 3.3x 10-4 mol phospholipid in 1 g of membrane 3.3x 10-4 mol phospholipid/ 1.5 x10-5 mol protein = 22 mol phospholipid/mol protein To estimate the percentage of the surface co ered by covered b phospholipid, phospholipid you o would o ld need to know (or estimate) the average crosssectional area of a phospholipid in a bilayer and the average crosssectional area of a 50 kDa protein