Key Final Fall 2009

Key Final Fall 2009 - lof3 BIS 102 Name K29 Fall 2009 Last...

Info icon This preview shows pages 1–4. Sign up to view the full content.

View Full Document Right Arrow Icon
Image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Image of page 2
Image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Image of page 4
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: lof3 BIS 102 Name K29 Fall, 2009 Last First K. Hilt Final Exam Score (200): Equations: pH = pK, + log {[b]/[a]} K, = XZ/(y-x) Kb = x2/ (y-x) pH = (pKall + pKaz)/2 (K,)(K.,) = 1 x 10'“ F=(q1q2)/sr2 AG=AH-TAS v,= {vm [S]}/ {Km+[S]} Amino acid pK.’s: a—carboxyl group (2.1), a—amino 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 pK.’s: C-terminal carboxyl group (3.6), N-terminal amino group (7.4) 1. (10 pts.) Liquid N2 is sometimes used to break open cells, in order to begin the purification of a protein. How does liquid N2 manage to break open cells? Explain briefly. Then give an example of when you would not want to use li'uid N2 and ex lain wh not. ‘ . ‘ 7435+ a+ Mq, can ‘tsp may“ Freezins “"145 W& ”1 firefi‘ M 2&1 H20; -o\e§-\—ro~3‘m a.” caMpMI-md'? 5"- ‘an. ct” . Dohi- use. it“ +e (”+42% av‘tmesd" \jm Wink ou evokin/en-ijfl I S I" 0“: cowafimaf'd F ‘50 a' 9 . 2. (30 pts.) Describe how you would purify hemoglobin fiom red blood cells. Assume that you start with 500 g of intact red blood cells. You are going to use an affinity column in your procedure. However, it the resin, run the column, and elute it. In all of your purification steps, you will need to run assays. What will they be? How will you eventually judge if your hemoglobin is pure? (Think about this). Your answer to all of the above items should show your mastery of several important topics that are relevant to protein purification. a, osmo'Hc assure. 41“?“ 5 y‘ ”am 5003 Cd‘s v.5“ byegk aye” ca“; I Cert ‘ 5% E “W P” 7.2—7.1. bet-HM k) “Hg 50 wash it saw . W5 «lea 2L ‘1 M * M ”06.5%” mgiim re 96 +__/—-—— f @m Qim, :41)?th . ‘ 39% a? :g" ‘If" A 54mg .. ‘3 O; b‘K’A‘I‘nS fisswj kslunbea in C‘AM.’ AA («robes/bk, Qu'e \(6 ‘ Q 0 z 1.7 Bra-a QDYA WWW“ 499”) ?“"e Ff 0 YID fiww More‘s; {n 0;_hs‘n&incj aukviinj/mj avoid“ “Wk. him QMR‘QLaA-‘tm 5W6. ’ 2) m )9ng - binmi pal-d m a. eerie: at m-Hve, 9%? 5d$ I‘L, I . J -' , . u.) +3 . ' rwz.‘ loanA m SDS- 9365' 'i "—11.5” WA) di?,:ubm;, 5 aitd 1 Land m 335": [75675343 at 6’7: 9L H-b, 3. (30 pts.) A stud needs to convert 2 liters of 150 mM phosphate buffer, pH 12.2 to a final pH of 2.5. How many moles of H+ will be necessary to achieve this? Assume that addition of the H+ does not attract the volume of the buffer. The pK.’s of phosphate are 2.12, 7.21, and 12.32. Put. your answer here: 5, I"! 2 mols of H". To receive credit, show all of your work on the back of this page. a- 5'. z L ' 3) I 0 q ( . 2 MIL 1+ ’ '5 We M H 5M*l (3” :2l ”5. 5:} 0'3?— 2J—l j 23 — (a? 0-3/2: '17- a» ._ ‘9. a. c.=‘ 334‘? 5 ,244 20f3 BIS 102 Name K % 4. (30 pts.) Explain the biochemistry of how C02 in animal tissues is removed by red blood cells, as the red blood cells pass from the tissues back to the lungs. Identify all of the key protein players. If biochemical reactions are involved, write them out as balanced Chemical expressions. Your overall anSWer Should he clear, thorough and cover the various aspects of the topic in depth. 0 x) CD; 4- play a: .—I @. swbumi‘rs 27’ “0-2, — l}; + (4+ A, PHD H 7.) C97, crosses REC Membrane. 0M3~ l5 cWaH—eh -\—o “£03 3 - k4, _ + ¢0z+ 147,0 carbonic“; we: ”C03 + H 3) HCD; and) Cl“ (ass +hrmjk Hg M‘Hemi-UI Hg; 2:90;... Mohave 5. (25 pts.) A particular “thiol protease” is known to have a catalytic triad that operates in a manner similar to that of chymotrypsin. What is different is that the thiol protease has a cysteine residue in place of the sen’ ' "he in chymoh'yps‘ in. The Other members of the catal" “ ’ ytic triad are the same. In the space below, draw the active site of the W showing the correct alignment, charges, and orientation of the catalytic triad members. Then indicate what each residue in the thiol protease catalytic triad is doing. (You do not need to draw in the substrate). If a residue is doing more than one thing, then list them all. ~. Jml; have+akwvfim /s+vW+0*“ ”" su 5 / I) ///\/\ f? fiwfl km ca‘h'an-fi' C 1' 6‘ C\ /c._ 047.“ [J l nudcofmle H V ong |) aria-«*6 wshxrc‘ shuelwwed— \ c1 2 3) (arms 5"i’“ 14-59ch «nus HeHAGAQ in cabk orien*a7\'\0V\ +mM55-Hw 5‘31;— 6. (20 pts.) Compound “ ” is a noncompetitive inhibitor of the thiol protease discussed in question #5 above. In the space below, draw the expected Lineweaver-Burk plots (with and without inhibitor), labeling theplotsandtheaxes"(axe‘sshouldhavéunitS).L' " "x- " ~13 ‘ ' ‘of ' "' 'lo .Thefiwrite'out the expected E + S :1. ES etc. expression describing the binding of the inhibitor. Lineweaver-Burk Plots + 1" E + S expression describing inhibition .1. E + 5 :2. E5 ——7 E- + p V0 + +- \ 1: I J. E1; + s 2;: E 3: VW __l_ T is} 3 of3 BIS 102 Name K 7. (10 pts.) Imagine that the Lineweaver-Burk plots that you drew in question #6 on the previous page were obtained using 30 uM enzyme in each assay. If you repeated each of the assays in those plots, but this time with juSt 15 uM enzyme in every assay, would the plots change? D6ferid your answer. Va. {7W will Lu, JA4‘4uevd'l bui— ‘J'Zm W‘H in... +9». same... / (20-06) 8. (30 pts.) What is the net charge, to the nearest 0.01, of the peptide -E-K-C-H-Y—T at pH 7.6? Put your answer here: -' 0 . '7 f . To receive credit, show yo wet} bfilo\\\ o 'l ,U 'WF' 4' H.“ W 01 PM" 01‘“ Hy" e“ : QKL+ ‘03, (a. . 7,1. '3 7- '4 + 103 ‘2 hi?“ 3.1M. 5330— (JAM o. z = 1,.) a W :59: b. . =4— - h ,.1__. a. .. “’ 2,52?’ ”He: 6.0 +19%} «- {0+4 W3“: 0.337 + (.0 4-03 + («0.lbb)[email protected])+(—IB r T ’F T 1‘ T n—ium. E K a .4 0mm. : “0.7S'I-(§ ‘= - o . 7f 9. (15 pts.) What is the final pH when 100 ml of 0.10 M ethanolamine is mixed with 150 ml of 0.20 M acetic acid? Put your answer here: pH = LI. 943 . The pKa’s of the protonated form of ethanolamine and acetic- aéid are 9.44 and 4.76, respectively. To receive credit, Show y‘éut‘ Wdl‘k Below. L0 . I an 9.30). I0 M) a 0x- 0! wine «Wflamiw (at 150,0.)(0,20 H) = 0503 we ext—OH" “3k 0 , 0‘ MA 043m“ read *9 awe. 0. a; run-Q $43600“ 0.0I mat/a‘zfl 9‘44 ‘5 ”‘76 + "1 Ab?— mre/n.zf,}\ : LL79 44»; 0’5 : LA71. +(‘a-30) 2 H.458? 2 ® ...
View Full Document

{[ snackBarMessage ]}

What students are saying

  • Left Quote Icon

    As a current student on this bumpy collegiate pathway, I stumbled upon Course Hero, where I can find study resources for nearly all my courses, get online help from tutors 24/7, and even share my old projects, papers, and lecture notes with other students.

    Student Picture

    Kiran Temple University Fox School of Business ‘17, Course Hero Intern

  • Left Quote Icon

    I cannot even describe how much Course Hero helped me this summer. It’s truly become something I can always rely on and help me. In the end, I was not only able to survive summer classes, but I was able to thrive thanks to Course Hero.

    Student Picture

    Dana University of Pennsylvania ‘17, Course Hero Intern

  • Left Quote Icon

    The ability to access any university’s resources through Course Hero proved invaluable in my case. I was behind on Tulane coursework and actually used UCLA’s materials to help me move forward and get everything together on time.

    Student Picture

    Jill Tulane University ‘16, Course Hero Intern