Solutions%20to%20Midterm

Solutions%20to%20Midterm - Solutions to Midterm 1a)...

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Solutions to Midterm 1a) Chymotrypsin cleaves on the carbonyl side of F, Y and W, producing the fragments: THEREDW INGSSTINK Staphylococcus aureus V8 protease cleaves on the carbonyl side of D and E, producing the fragments: THE RE D W INGSSTINK 1b) The pI for fragment D is 2.77 and fragment W is 5.89 (Table 3-1 page 78). The other fragments require pI to be calculated: Fragment THE pH N-terminal of T (9.62) C-terminal of E (2.19) Side Chain of H (6.00) Side Chain of E (4.25) Net Charge 1 + 0 + 0 +2 3 + - + 0 +1 5 + - + - 0 7 + - 0 - -1 pI = 0.5(4.25 + 6.00) = 5.13 Fragment RE pH N-terminal of R (9.04) C-terminal of E (2.19) Side Chain of R (12.48) Side Chain of E (4.25) Net Charge 1 + 0 + 0 +2 3 + - + 0 +1 5 + - + - 0 10 0 - + - -1 pI = 0.5(4.25 + 9.04) = 6.65
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Fragment INGSSTINK pH N-terminal of I (9.68) C-terminal of K (2.18) Side Chain of K (10.53) Net Charge 1 + 0 + +2 3 + - + +1 10 0 - + 0 11 0 - 0 -1 pI = 0.5(9.68 + 10.53) = 10.1 1c) At a pH of 7.0, all fragments with a pI lower than 7.0 will be negatively charged and will be bound to an anion exchanger. Thus only INGSSTINK will not be retained on the anion exchanger. 2a) Pepsin cleaves on the amino of F, Y and W, producing the fragments: SENS, WILLAL, WA, YSDE, FEATTHEMAPLELEA, FS Chymotrypsin cleaves on the carbonyl side of F, Y and W, producing the fragments: SENS, W, ILLAL, W, A, Y, SDE, F, EATTHEMAPLELEA, F, S 2b) The average molecular mass of an amino acid residue is 110 Da, thus: SENS: 440 Da ILLAL: 550 Da SDE: 330 Da EATTHEMAPLELEA: 1540 Da All single amino acids weigh 110 Da 2c) EATTHEMAPLELEA will elute first, followed by ILLAL and then by SENS. SDE and all the single amino acids are below the range of the column and thus will not separate. They will all co-elute together after the other fragments have eluted. FYI: I have no bias towards any hockey team what so ever….
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3) Solution A contains 0.25 M Ca(OH) 2 and 1.75 M sucrose. Solution B contains 0.50 M Na 2 S and 0.50 M Ca(CO 3 ). The osmolarity of solution A is: 3(0.25) + 1(1.75) = 2.50
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Solutions%20to%20Midterm - Solutions to Midterm 1a)...

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