Biochemistry
9th Edition
ISBN: 9781305961135
Author: Mary K. Campbell, Shawn O. Farrell, Owen M. McDougal
Publisher: Cengage Learning
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Chapter 5, Problem 22RE
RECALL How does HPLC differ from ion-exchange chromatography?
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The beta-lactamase hydrolyzes the lactam-ring in penicillin. Describe the mechanism
of hydrolysis, insuring to include the involvement of S, D, & K in the reaction sequence. Please help
To map the active site of beta-lactamase, the enzyme was hydrolyzed with trypsin to yield a hexapeptide (P1) with the following amino acids. Glu, Lys, Leu, Phe, Met, and Ser. Treatment of P1 with phenyl isothiocyanate yielded a PTH derivative of phenylalanine and a peptide (P2). Treatment of P1 with cyanogenbromide gave an acidic tetrapeptide (P3) and a dipeptide (P4).Treatment of P2 with 1-fluoro-2,4-dinitrobenzene, followed by complete hydrolysis, yields N-2,4-dinitrophenyl-Glu. P1, P2, and P3 contain the active site serine.
Why doesn't D in this hexapeptide not participate in the hydrolysis of the beta-lactam ring even though S, K, and D are involved in the catalyst?
To map the active site of -lactamase, the enzyme was hydrolyzed with trypsin to yield a hexapeptide (P1) with the following amino acids. Glu, Lys, Leu, Phe, Met, and Ser. Treatment of P1 with phenyl isothiocyanate yielded a PTH derivative of phenylalanine and a peptide (P2). Treatment of P1 with cyanogenbromide gave an acidic tetrapeptide (P3) and a dipeptide (P4).Treatment of P2 with 1-fluoro-2,4-dinitrobenzene, followed by complete hydrolysis, yields N-2,4-dinitrophenyl-Glu. P1, P2, and P3 contain the active site serine.
Using the experimental results described above derive the primary sequence of the active site hexapeptide. Please help!
Chapter 5 Solutions
Biochemistry
Ch. 5 - RECALL What types of homogenization techniques are...Ch. 5 - RECALL When would you choose to use a...Ch. 5 - RECALL What is meant by salting out? How does it...Ch. 5 - RECALL What differences between proteins are...Ch. 5 - RECALL How could you isolate mitochondria from...Ch. 5 - RECALL Can you separate mitochondria from...Ch. 5 - RECALL Give an example of a scenario in which you...Ch. 5 - Prob. 8RECh. 5 - REFLECT AND APPLY You are purifying a protein for...Ch. 5 - Prob. 10RE
Ch. 5 - RECALL What is the basis for the separation of...Ch. 5 - RECALL What is the order of elution of proteins on...Ch. 5 - RECALL What are two ways that a compound can be...Ch. 5 - Prob. 14RECh. 5 - RECALL Why do most people elute bound proteins...Ch. 5 - RECALL What are two types of compounds that make...Ch. 5 - RECALL Draw an example of a compound that would...Ch. 5 - RECALL How can gel-filtration chromatography be...Ch. 5 - REFLECT AND APPLY Sephadex G-75 has an exclusion...Ch. 5 - Prob. 20RECh. 5 - RECALL What is the main difference between reverse...Ch. 5 - RECALL How does HPLC differ from ion-exchange...Ch. 5 - REFLECT AND APPLY Design an experiment to purify...Ch. 5 - REFLECT AND APPLY Referring to Question 23, how...Ch. 5 - Prob. 25RECh. 5 - REFLECT AND APPLY You wish to separate and purify...Ch. 5 - REFLECT AND APPLY An amino acid mixture consisting...Ch. 5 - REFLECT AND APPLY An amino acid mixture consisting...Ch. 5 - REFLECT AND APPLY In reverse-phase HPLC, the...Ch. 5 - REFLECT AND APPLY Gel-filtration chromatography is...Ch. 5 - RECALL What physical parameters of a protein...Ch. 5 - RECALL What types of compounds make up the gels...Ch. 5 - RECALL Of the two principal polymers used in...Ch. 5 - RECALL What types of macromolecules are usually...Ch. 5 - RECALL If you had a mixture of proteins with...Ch. 5 - RECALL What does SDSPAGE stand for? What is the...Ch. 5 - RECALL How does the addition of sodium...Ch. 5 - RECALL Why is the order of separation based on...Ch. 5 - RECALL The accompanying figure is from an...Ch. 5 - Prob. 40RECh. 5 - Prob. 41RECh. 5 - Prob. 42RECh. 5 - Prob. 43RECh. 5 - REFLECT AND APPLY What would happen during an...Ch. 5 - REFLECT AND APPLY A sample of an unknown peptide...Ch. 5 - REFLECT AND APPLY A sample of a peptide of unknown...Ch. 5 - REFLECT AND APPLY You are in the process of...Ch. 5 - REFLECT AND APPLY You are in the process of...Ch. 5 - Prob. 49RECh. 5 - Prob. 50RECh. 5 - Prob. 51RECh. 5 - Prob. 52RECh. 5 - Prob. 53RECh. 5 - Prob. 54RECh. 5 - RECALL What is the basis for the technique called...Ch. 5 - Prob. 56RECh. 5 - Prob. 57RECh. 5 - RECALL What are the main procedures involved in a...Ch. 5 - RECALL Where did western blot get its name?Ch. 5 - Prob. 60RECh. 5 - Prob. 61RECh. 5 - Prob. 62RECh. 5 - Prob. 63RECh. 5 - Prob. 64RECh. 5 - Prob. 65RE
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