Enzymology: enzymes

Chapters 7-9 Emphasis on basic enzymology— enzyme kinetics, mechanisms and regulation. 

Coleman, D. E., and Sprang, S. R. (1999a). Reaction dynamics of G-protein catalyzed hydrolysis of GTP as viewed by X-ray crystallographic snapshots of Gial. In Methods in Enzymology Enzyme Kinetics and Mechanism, Part E Energetics of Enzyme Catalysis (V. L. Schramm and D. L. Puvich, Eds.), Vol. 308, pp. 70-92. Academic Press, New York. 

Price, N. C. and Stevens, L. (1999). Fundamentals of Enzymology Enzyme turnover (Third edition, pp.370-399). Great Clarendon, Oxford Oxford University Press. 

Methods in Enzymology, Enzyme Structure, Vol. 91, Academic Press, New York... 

Principles of Clinical Enzymology Enzymes Enzymes Appendix... 

Pace, C. N. 1986. Determination and analysis of urea and guanidine hydrochloride denatur-ation curves. In Methods in Enzymology. Enzyme Structure Part L, edited hy C. H. W. Hits and S. N. Timasheff. Orlando, FL Academie Press, p. 266. 

The high sensitivity of fluorescence spectroscopy and the selectivity of enzymatic assays are responsible for the increasing use of fluorimetric methods in enzymology. Enzyme determinations usually involve the use of kinetic methodology for measuring the rate of formation of the fluorescent product, while both equilibrium and kinetic methods are used to determine the substrates. Fluorimetric measurements on enzyme-catalyzed reactions have been used for a long time to determine a variety of enzymes and substrates (Figure 2). 

Scheindlin, Stanley. Clinical Enzymology Enzymes As Medicine. Molecular Interventions 7, no. 1... 

Brand, L., and B. Witholt Fluorescence Measurements, in Methods in Enzymology-Enzyme Structure (C. H. W. Hirs, Ed.). 11,776-856. London-New York Academic Press. 1973. 

Perhaps the biggest impact on the practical utilization of enzymes has been the development of nonaqueous enzymology (11,16,33,35). The use of enzymes in nonaqueous media gready expands the scope of suitable transformations, simplifies thek use, and enhances stabiUty. It also provides an easy means of regulation of the substrate specificity and regio- and enantioselectivity of enzymes by changing the reaction medium. 

This chapter lists some representative examples of biochemicals and their origins, a brief indication of key techniques used in their purification, and literature references where further details may be found. Simpler low molecular weight compounds, particularly those that may have been prepared by chemical syntheses, e.g. acetic acid, glycine, will be found in Chapter 4. Only a small number of enzymes and proteins are included because of space limitations. The purification of some of the ones that have been included has been described only briefly. The reader is referred to comprehensive texts such as the Methods Enzymol (Academic Press) series which currently runs to more than 344 volumes and The Enzymes (3rd Edn, Academic Press) which runs to 22 volumes for methods of preparation and purification of proteins and enzymes. Leading referenees on proteins will be found in Advances in Protein Chemistry (59 volumes. Academic Press) and on enzymes will be found in Advances in Enzymology (72 volumes, then became Advances in Enzymology and Related Area of Molecular Biology, J Wiley Sons). The Annual Review of Biochemistry (Annual Review Inc. Patio Alto California) also is an excellent source of key references to the up-to-date information on known and new natural compounds, from small molecules, e.g. enzyme cofactors to proteins and nucleic acids. 

T. C. Bruice and S. I Benkovic, Bioorganic Mechanisms, Vol. 1, W. A. Benjamin, New brk, 1966, pp. 1-258 W. P. Jencks, Catalysis in Chemistry and Enzymology, McGraw-Hill, New York, 1969 M. L. Bender, Mechanisms of Homogeneous Catalysis from Protons to Proteins, Wiley-Interscience, New York, 1971 C. Walsh, Enzymatic Reaction Mechanisms, W. H. Freeman, San Francisco, 1979 A. Fersht, Enzyme Structure and Mechanism, 2nd ed., W. H. Freeman, New York, 1985. 

Wn, R., 1993. Development of enzyme-ba.sed mediods for DNA. sequence analy.sis and dieir application in genome projects. Methods in Enzymology 67 431-468. 

Silverman, R. B., 1988. Mechanism-Based Enzyme Inactivation Chemistry and Enzymology, Vols. I and II. Boca Raton, FL CRC Press. 

Enzymology of proteases in a water-phase is well known, but its alteration in a compartment is poorly understood. There are dramatical changes in reaction rates, in enzyme contractions and in enzyme sensitivity to inhibitors, which are not exactly described. In addition, besides fibrin and platelets there are several cellular and molecular components present in a thrombus compartment, where their influence on the basic fibrinolytic reactions is not known. To study this aspect of fibrinolysis is a task of the near future [4]. 

S. P. Colowick u. N. O. Kaplan, Methods in Enzymology, Vol. XXII, Enzyme Purification and Related Techni-... 

K. Mosbach. Methods in Enzymology XLIV Immobilized Enzymes, Academic Press, New York 1976. 

Plasma also contains numerous other enzymes that perform no known physiologic function in blood. These apparently nonfunctional plasma enzymes arise from the routine normal destruction of erythrocytes, leukocytes, and other cells. Tissue damage or necrosis resulting from injury or disease is generally accompanied by increases in the levels of several nonfunctional plasma enzymes. Table 7-2 lists several enzymes used in diagnostic enzymology. 

Schroeder, W. A. "Separation of Peptides by Chromatography on Columns of Dowex 1 with Volatile Developers", In "Methods In Enzymology", p. 214, Vol. XXV, "Enzyme Structure, Part B", C. H. W. Hlrs and S. N. Tlmasheff, Editors, Academic Press, New York, 1972. 

It has been demonstrated that such controls are not valid and that there may be significant differences in enzyme concentrations between two cell types (56). For this reason, normal amniotic fluid cells themselves must be used as controls for amniotic fluid cell cultures being subjected to enzymologic inves t igat ion. 

A Pstl-HinA (1.2 kbp) fragment was subcloned in pUC19. The enzyme produced by the E. coli transformant was purified fo homogeneify and shown to be identical to that of the original strain. Both enzymes had the same enzymological properties and N-terminal amino acid sequences. ... 

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