Laboratory Guide to Biochemistry, Enzymology, and Protein Physical Chemistry

1. Front Matter

   Pages i-xii

   PDF

2. Aspartate Transcarbamylase

   • Marc le Maire, Raymond Chabaud, Guy Hervé

   Pages 1-15

3. Molecular Genetics

   • Marc le Maire, Raymond Chabaud, Guy Hervé

   Pages 17-24

4. Purification of Aspartate Transcarbamylase and Its Subunits

   • Marc le Maire, Raymond Chabaud, Guy Hervé

   Pages 25-32

5. Structural and Physicochemical Study of Aspartate Transcarbamylase

   • Marc le Maire, Raymond Chabaud, Guy Hervé

   Pages 33-89

6. Enzymatic Catalysis and Regulation

   • Marc le Maire, Raymond Chabaud, Guy Hervé

   Pages 91-140

7. Complementary Experiments

   • Marc le Maire, Raymond Chabaud, Guy Hervé

   Pages 141-142

8. Back Matter

   Pages 143-169

   PDF

The study of a single well-chosen substance, here aspartate transcarb­ amylase, can provide an excellent basis for a laboratory course. The student is introduced to a variety of scientific ideas and to many experi­ mental and interpretive techniques. This enzyme is readily available, is relatively stable, has an extensive literature, and its behavior has many facets: substrate inhibition, a large change in structure upon homo­ tropic activation by substrates, allosteric stimulation by ATP, allosteric inhibition by CTP synergistic with VTP, positive cooperativity for sub­ strates, negative cooperativity for CTP binding, and dissociation and reassembly of subunits Cand R2 from the holoenzyme CI\5. In addition 3 6 to the known biochemical aspects of these properties, the results ob­ tained here can be interpreted in the light of the high-resolution X-ray diffraction structures of the T and R forms, the low-angle X-ray scattering results, and the large number of mutants now available by recombinant DNA methods. Future development of this course could also involve part of these methods, as well as the carefully chosen experiments described here. This approach resembles research more than the approaches one usually finds in biochemical laboratory courses. A consistent develop­ ment of ideas about a single enzyme, which shows so many facets in its behavior, is sure to hold the interest of the student. Moreover, one explores a depth, and reasons to move forward, that are an essential part of research.

• enzyme
• enzymology
• physical chemistry
• structure

• Centre National de la Recherche Scientifique, France

  Marc Maire, Guy Hervé

• Université Pierre et Marie Curie, Paris, France

  Raymond Chabaud

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