ANALYTICAL ENZYMOLOGY

Analytically the clinical laboratorian is concerned with measuring the activity or mass in serum or plasma of enzymes that are predommantly intracellular and that are normally present in the serum in low levels only. By measuring changes in the levels of these enzymes in disease, it is possible to infer the location and nature of pathological changes m the tissues of the body. 

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Enzymes are proteins with catalytic properties clinical enzymology is the application of the science of enzymes to the diagnosis and treatment of disease. The principles of clinical enzymology will be mtroduced and discussed in this chapter. Individual topics include basic principles, enzyme kinetics, analytical enzymology, and diagnostic enzymology. 

Fields, G.B. (Ed.) (1997) Methods in Enzymology. Vol 289. Solid-phase Peptide Synthesis. 710pp. Academic Press. Includes analytical techniques. 

D Holme and H Peck, Analytical Biochemtstiy, 3rd ed (1998), Addison Wi li Longman (New York), pp 406-442 Structure, function, and analysis of lipids C Matthews, K van Holde, and K Ahern, Biochemistry, 3rd ed (2000), Benjamin/ Cummings (San Francisco), pp 315-357 Lipid structure and function G Patton, S Cann, H Brunengraber, and J. Lowenstein, in Methods in Enzymology, Vol 72, J Lowenstein, Editor (1981), Academic Press (New York), pp 8-20 Separation of fatty acid methyl esters by gas chromatography on capillary columns N Radin, in Methods in Enzymology, Vol 72, J M Lowenstein, Editor (1981), Academic Press (New York), pp 5-7 Extraction of lipids with hexane-isopropanol L Stryer, Biochemistry, 4th ed (1995), W H Freeman (New York), pp 263-270, 603-606 Lipid structure and function... 

Work on dithiolene chemistry continued through the 1960s and 1970s fueled by continued interest in the remarkable coordination chemistry of dithiolene complexes. However, in the last 20 years tremendous added impetus to research in the area arose from discoveries in materials science, enzymology, analytical science, and reactivity that broadened the impact and import of dithiolene chemistry. This volume seeks to capture the interplay of basic work on dithiolene complexes with the growing biological, sensor, reactivity, and materials science implications and applications that have made dithiolene chemistry a vibrant and growing held. 

Labeled proteins are used in enzymology especially as substrates for determining the activity of proteolytic enzymes, for studying structural properties of enzyme molecules (7.2), and for some analytical purposes. Here, attention is paid to the determination of proteolytic activities when proteins labeled with radionuclides, chromophores and fluorophores are used as substrates. 

As an analytical spectroscopic technique, EPR is similar in concept to the more widely used nuclear magnetic resonance (NMR) spectroscopy [see NMR Overview of Applications in Chemical Biology]. In fact, EPR and NMR are complementary to each other. Both techniques detect magnetic moments, hut NMR determines the chemical stmctures in solution, whereas EPR describes more precisely the electronic and chemical structures of a particular region of the biological system, such as electron transfer centers, metal ions, and an intermediate state of the enzyme or substrate. It is not possible to present a full description of the theory of EPR in an article with this scope. Therefore, only sufficient information is provided here to enable the readers to understand the practical aspects of this analytical tool in enzymology. 

McComb RB. The measurement of lactate dehydrogenase. In Homburger HA, ed. Clinical and analytical concepts in enzymology. Skokie, IL College of American Pathologists, 1983 157-71. 

This view of the living organism as a complex network of enzyme-catalyzed reactions derives in large part from the well established analytical tradition in biochemistry known as enzymology. 

Analytical techniques applicable to steroids and the synthesis of isotopically labelled steroids of high specific activity are amongst the topics reviewed in the latest volume of a comprehensive treatise on enzymology and the first volume of a new series of books on steroid biochemistry includes a review of the steroids found in marine invertebrates and plants. 

DeLuca, M. A., ed., Methods in Enzymology, Vol. 57. Academic Press, New York, 1978. DeLuca, M, A., and McElroy, W. D., eds., Bioluminescence and Chemiluminescence Basic Chemistry and Analytical Applications (Proceedings of the Second International Symposium on Bioluminescence and Chemiluminescence) Academic Press, New York, 1981. 

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