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Final Exam with Answer Key for Structure Function Biomolecules | BIS 102, Exams of Biology

Material Type: Exam; Professor: Hilt; Class: Struc-Func Biomolecules; Subject: Biological Sciences; University: University of California - Davis; Term: Fall 2012;

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lof4 BIS 102 Name Key Fall, 2012 Last 7 First K. Hilt Final Exam Score (200): Equations: pH = pK, + log {[bVa}} AG = AH - TAS Vo = {Vmax [S]}/ {Km + [S]} K,=x'Ay-x) Kp=x/(y-x) pH =(Ka, + pKa,)/2 pK,’s of amino acid side chains: _D (3.9), E (4.2), H (6.0), C (8.3), ¥ (10.1), K (10.5), R (12.5) pK, of amino acid a-COOH (2.1); pK, of amino acid o-NH;* (9.6) pK, of oligopeptide N-terminus (7.4); pK, of oligopeptide C-terminus (3.6) 1, Enzyme Z catalyzes the reaction; A -+NAD* <> B+NADH + H* Besides following the [H'] using a pH meter, the reaction can be monitored spectroscopically at 340 nm. This is the wavelength at which NADH absorbs light. Some of the first enzymes that were worked on in biochemistry were dehydrogenases, like our enzyme Z, because of this advantage. Radioisotopes were not yet available. (5 pts.) a) Diagram what the kinetic assay results would be if the reaction was measured from right to left. Use the axes drawn below. Label the axes. {weioH] Fra40 Time (15 pts.) b) Describe in detail how you would accurately determine enzyme Z's Km’s for NAD* and A. Describe the experimental set-up and the analysis of data. What is going into each cuvette? What is in excess? What is being varied? What is the same? What are you plotting? What are you reading off of the me For Ne + pub in egcena A Pived evnceahation of. enrgme & ond beer anh very [witot). Meke a bineweaver — Burk ple Cho vs. “Vineev)) and x-tnlercaptt = -%,. Zz. For A: peck im evcese [Wart] | same Syed comatwition of entymet and bu as obwe, wut vavy CA]. Make a Lineweaver~ Buk let (Ap us. Vial) andl xv intercept = (15 pts.) c) Imagine that twenty I.U.’s of enzyme Z were catalyzing the above reaction for one minute, under Vax conditions, in a 3.00 ml assay volume. The assay is buffered with 20 mM phosphate buffer, pH 7.80, What will the pH be at the end of that one minute? Put your answer here: final pH=_7./4 . To receive credit, show all of your work on the back of this page. The pK, ’s of H3PO, are 2.12, 7.21, and 12,32,