BIS102 Hilt SSII07 FINAL

BIS102 Hilt SSII07 FINAL - lof2 BIS 102 Name IQ? Summer,...

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Unformatted text preview: lof2 BIS 102 Name IQ? Summer, 2007 Last First K. Hilt Final Exam Score (200): Equations: pH = pKa + log {[b]/[a]} K, = xz/(y—x) Kb = x2/ (y-x) pH = (pKal + pKa2)/2 (K,)(Kb) = 1 x 10'” F=(qlq2)/sr2 AG=AH-TAS v.,= {vm[s1}/ {Km+[S]} Amino acid pKa’s: a—carboxyl group (2.1), a-arnino group (9.6) side chains: D (3.9) E (4.2) H (6.0) C (8.3) Y (10.1) K (10.5) R (12.5) Oligopeptide pKa’s: C-terminal carboxyl group (3.6), N-tenninal amino group (7.4) 1. A Michaelis—Menten type enzyme gave an initial velocity of 25 umol P in 2 min., when assayed at an initial substrate concentration of 34 uM, which happened to be the Km of the enzyme. The MW of the enzyme is 65,000 g/mol. (10 pts.) a) What is Vmax? Put your answer on this line (include units): 5‘0 EMA [2 min . To receive credit, you must show your work on the back of this page. There is no artial credit. Ku': Va, Vina/5‘ *‘- VMW : zszflWefflwk3= S'O/A-MOQP/Zmin. (10 pts.) b) What would v() be if the same amount of enzyme was assayed at a substrate concentration of 15 uM? Put your answer on this line (include units): I 5‘. 3 £1 mreZZ min . To receive credit, you must show your work on the back of this a c. There is no partial credit. . a we»; is ,_ (rofiMf/zmk 15' H t so/me/zw‘fi 05' ) a We ‘ W W “’fi'af—Ti—A "*3/“ AW“- (15 pts.) c) If e above assays wer each carried out in 3.00 ml volumes’ffe. total volume of the assay is 3.00 ml), and each assay contained 4.25 pg of pure enzyme, what is the turnover number of this enzyme? Put your answer on this line (include units): 3 .9 x w ‘- miu’ ' . To receive credit, you must show your work on the back. of this age. There is no partial credit. : 0 me = 2, "We ' _ 25x11)" p./"‘“‘ I = ‘ 5— 2. (40 pts.) Calculate the missing values (X)'in the following enzyme purification table: 3 W0 5; M. Jr 6 Volume [Protein] Total Activity Specific Fraction (ml) m 1 ml) I.U./ml I.U. Yield (%) Activi Purification .15- 2. resuspended 200 20 X2 X3 85 X4 X5 40;? (“102504 an; 2550 4.6375 3.12 o ‘ Ct . 3. afi‘mi column I Lu. You do not have to show your calculations. %?/f 3. (15 pts.) What is the basis of protein migration in each of the following gels? Your answer is restricted to just the space on each line. If a protein migrates due to more than one factor, then list them all. However, do not guess. Additional wrong answers make the whole answer incorrect. 4’; a) SDS-PAGE gel size. (ma/V1. +5 b) IEF gel 1: +1; c) Native PAGE gel lm MAL size 4. (10 pts.) Why is a stacking gel used in native PAGE gels? Restrict your answer to the space on these two lines: ‘m creed); bani «e sow-H ‘k. mvm 2of2 5. (20 pts.) How many moles of H+ must be added to 250 ml of 0.100 M histidine buffer, pH 10.1, to change the pH to 2.2? Show all calculations. Assume that the addition of H+ does not affect the total buffer volume. Put your answer on this line: 0 . 0 S r va . To receive credit, you must show your work on the back of page 1. There is no partial credit. 6. (20 pts.) What is the pH of a 100 ml 0.250 M ammonia buffer whose net charge is “+0.83”? Show all calculations. Note: the Kb of NH3 is 1.8 x 10's. Show all calculations in the space below. There is no partial credit. -gq (imam = W ‘0 m, a.” LG Ka. -=. 1‘10 __§=§';$’ 1.8/x10 95’”; 04"" ‘W% 6.17 = «.29 + [0} 03,3 : mu. + (-0.04) a 7. (30 pts.) Amino acid analysis of protein “Z” gave the following experimental results: in Protein “Z” in Protein “Z” _ G — [on-mm. I . c-l-crm.—l “PM -;~. 37 + What is the net charge (to the nearest 0.01) of protein “Z” at pH 8.4? Put your answer on the following “J65. line: 27. 3-} . To receive credit, you must show all of your work on the back of this page. 8. 30 ts. Fill in the followin fibula Positive or Binding Site(s) Strengthen, Create, and/or x 7, Ne ative? Break Salt Brid es? z'ii‘:fi:.:.%‘::i“~ “at. . an... C02 “ejaz’vt N_,K_VM;“; a", a“ subunfl'j 5. gig(o.z;o£,)(o.w M) = 0, our M 1+ lit/“W 341 2.! +1 6‘0 0 «.0 fl H”; ‘2—— H :1... H 7;_ H T .& e’r\'=|Dl ‘ 2‘2 ‘7 zzzztfipwkh w.z—;qa+/,}£ &. 0.5-; 0.\ " 3,15 —.-_. k; u. b: 3.”, T; 0-7; [.20 => 1 ' I hub “7‘ *7”? "z 0- t. 015M ".2 0, 0,4 M + .AngEgpqq L > H “clue Lo.uq)(o.:z§m£)= 0.0,, We w. m 2 l rd fi+W=T &,0U f— 0.0;; +o,olf z 0. 05‘5‘ \ Mmeci—ikukm7ke 9H : ’90,. 4- “"1 7 "L luj/% Ck *— K PL h e“ r» a. 9»! =93+T§gfi 0.: 7. lag}: . .. . (“0.59m :— :UfiM 9‘“va "' 37 *‘(0-0‘1)+C—a.227) 4—09.52» 1 27.3% ...
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This note was uploaded on 10/07/2009 for the course BIS 52192 taught by Professor Enochbaldwin during the Summer '09 term at UC Davis.

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BIS102 Hilt SSII07 FINAL - lof2 BIS 102 Name IQ? Summer,...

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