Biochemistry, The Molecular Basis of Life, 6th Edition
6th Edition
ISBN: 9780190259204
Author: Trudy McKee, James R. McKee
Publisher: Oxford University Press
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Chapter 5, Problem 25RQ
Summary Introduction
To review:
The types of chromatography involved in the purification of proteins and the description of these methods.
Introduction:
Chromatography is a technique for separation of proteins present in a mixture. Proteins separated on the basis of their shape, size, weight, and binding affinities. Chromatography involves two phases, a mobile phase (generally liquid), during which the protein mixture is dissolved, and a stationary phase, during which the protein travels through a solid matrix.
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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, The Molecular Basis of Life, 6th Edition
Ch. 5 - Prob. 1QCh. 5 - Prob. 2QCh. 5 - Prob. 3QCh. 5 - Prob. 4QCh. 5 - Prob. 5QCh. 5 - Prob. 6QCh. 5 - Prob. 7QCh. 5 - Prob. 8QCh. 5 - Prob. 9QCh. 5 - Prob. 10Q
Ch. 5 - Prob. 11QCh. 5 - Prob. 12QCh. 5 - Prob. 1RQCh. 5 - Prob. 2RQCh. 5 - Prob. 3RQCh. 5 - Prob. 4RQCh. 5 - Prob. 5RQCh. 5 - Prob. 6RQCh. 5 - Prob. 7RQCh. 5 - Prob. 8RQCh. 5 - Prob. 9RQCh. 5 - Prob. 10RQCh. 5 - Prob. 11RQCh. 5 - Prob. 12RQCh. 5 - Prob. 13RQCh. 5 - Prob. 14RQCh. 5 - Prob. 15RQCh. 5 - Prob. 16RQCh. 5 - Prob. 17RQCh. 5 - Prob. 18RQCh. 5 - Prob. 19RQCh. 5 - Prob. 20RQCh. 5 - Prob. 21RQCh. 5 - Prob. 22RQCh. 5 - Prob. 23RQCh. 5 - Prob. 24RQCh. 5 - Prob. 25RQCh. 5 - Prob. 26RQCh. 5 - Prob. 27RQCh. 5 - Prob. 28RQCh. 5 - Prob. 29RQCh. 5 - Prob. 30RQCh. 5 - Prob. 31RQCh. 5 - Prob. 32FBCh. 5 - Prob. 33FBCh. 5 - Prob. 34FBCh. 5 - Prob. 35FBCh. 5 - Prob. 36FBCh. 5 - Prob. 37FBCh. 5 - Prob. 38FBCh. 5 - Prob. 39FBCh. 5 - Prob. 40FBCh. 5 - Prob. 41FBCh. 5 - Prob. 42SACh. 5 - Prob. 43SACh. 5 - Prob. 44SACh. 5 - Prob. 45SACh. 5 - Prob. 46SACh. 5 - Prob. 47TQCh. 5 - Prob. 48TQCh. 5 - Prob. 49TQCh. 5 - Prob. 50TQCh. 5 - Prob. 51TQCh. 5 - Prob. 52TQCh. 5 - Prob. 53TQCh. 5 - Prob. 54TQCh. 5 - Prob. 55TQCh. 5 - Prob. 56TQCh. 5 - Prob. 57TQCh. 5 - Prob. 58TQCh. 5 - Prob. 59TQCh. 5 - Prob. 60TQCh. 5 - Prob. 61TQCh. 5 - Prob. 62TQCh. 5 - Prob. 63TQCh. 5 - Prob. 64TQCh. 5 - Prob. 65TQCh. 5 - Prob. 66TQCh. 5 - Prob. 67TQCh. 5 - Prob. 68TQCh. 5 - Prob. 69TQCh. 5 - Prob. 70TQ
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