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Extraction and Assay of Ornithine Decarboxylase: A Laboratory Experiment that Introduces Principles of Radiochemical Enzymatic Assay

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William H. Voige

Department of Chemistry, James Madison University, Harrisonburg, VA 22807

J. Chem. Educ., 1997, 74 (8), p 988

DOI: 10.1021/ed074p988

Publication Date (Web): August 1, 1997

Section:

History, Education, and Documentation

Abstract

A laboratory experiment involving preparation of an Escherichia coli extract in which ornithine decarboxylase (ODC) is quantitated by radiochemical assay has been developed. The assay is based on the ODC-catalyzed decarboxylation of [1-¹⁴C]ornithine to yield ¹⁴CO₂. Procedures for growth of the bacteria, preparation of the extract and assay of the enzyme are described. Points to be considered before adopting the experiment and potential problems are discussed. The experiment gives students experience in laboratory procedures, safety precautions and calculations associated with the use of radioactive compounds. It introduces principles common in radiochemical work and illustrates sensitivity and selectivity of a well-designed enzyme assay. It is often the first time students encounter a situation in which enzyme stereospecificity must be taken into account in a laboratory procedure. The experiment is suitable for use in biochemistry, radiochemistry or chemically-oriented cell biology laboratory classes.

Keywords (Audience):

Upper-Division Undergraduate

Keywords (Domain):

Biochemistry

Keywords (Pedagogy):

Hands-On Learning / Manipulatives

Keywords (Subject):

Enzymes

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This article has been cited by 1 ACS Journal articles (1 most recent appear below).

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Journal of Chemical Education2006 83 (2), 294
- The Binding Constant of Estradiol to Bovine Serum Albumin. An Upper-Level Experiment Utilizing Tritium-Labeled Estradiol and Liquid Scintillation Counting
  Peihong Liang , Bhavin Adhyaru , Wright L. Pearson and Kathryn R. Williams
  Journal of Chemical Education2006 83 (2), 294
  Students determine the binding constant of estradiol to bovine serum albumin using 3H-labeled estradiol. The estradiol is equilibrated with excess BSA, and subsequently a small fraction (ca. 1%) of the unbound hormone is extracted using solid-phase ...
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