Advertisement

The Study of Fast Reactions by the Stopped Flow Method

  • Wolfgang H. Goldmann
  • Zeno Guttenberg
  • Robert M. Ezzell
  • Gerhard Isenberg
Part of the Principles and Practice book series (PRINCIPLES)

Abstract

The mechanisms involved in protein reactions may be elucidated by investigating the reaction kinetics. There are two investigative approaches in ascertaining the combination of elementary steps which constitute these mechanisms:(1) steady state kinetics permits analysis of the overall reaction in which protein substrates are converted into products without examining the protein molecule itself; and (2) transient kinetics allows the direct measurement of each component in the overall reaction. In this latter case, attention is focused on changes occurring in the molecule upon binding to another protein whereas the study of the equilibrium (steady state) reaction does not examine the protein molecule directly. Equilibrium studies have wider applicability since they usually require only a small amount of protein and do not involve the use of special equipment. Necessarily, the information obtained is indirect and often ambiguous. Although transient kinetics requires special techniques for measuring the rates of fast reactions in solutions, it provides information which is far more direct and useful for elucidating complicated mechanisms of reactions. Thus, the two approaches are complementary and both are indispensable for the study of protein reactions.

Keywords

Fast Reaction Protein Reaction Elementary Step Steady State Kinetic Stopped Flow 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bernasconi CF (1976) Relaxation kinetics. Academic Press, New YorkGoogle Scholar
  2. Burridge K, Connell L (1983) A new protein of adhesion plaques and ruffling membranes. J Cell Biol 97:359–367PubMedCrossRefGoogle Scholar
  3. Craig SW, Lancashire CL, Cooper JA (1982) Methods Enzymol 85:316–335PubMedCrossRefGoogle Scholar
  4. Detmers P, Weber A, Elzinga M, Stephens RE (1981) 7-Chloro-4-nitrobenzeno-2-oxa-l,3-diazole actin as a probe for actin polymerization. J Biol Chem 256:99–10PubMedGoogle Scholar
  5. Eccleston JF (1987) Spectrophotometry and spectrofluorimetry:a practical approach, chap 6. In:Bashford CL, Harris DA (eds) Stopped-flow spectrophotometric techniques. IRL Press, OxfordGoogle Scholar
  6. Geiger B (1979) A 130 K protein from chicken gizzard:its localization at the termini of microfilament bundles in cultured chicken cells. Cell 18:193–205PubMedCrossRefGoogle Scholar
  7. Goldmann WH, Geeves MA (1991) A “slow” temperature jump apparatus built from a stopped-flow machine. Anal Biochem 192:55–58PubMedCrossRefGoogle Scholar
  8. Gutfreund H (1972) Enzymes:physical principles. Wiley-Interscience, LondonGoogle Scholar
  9. Gutfreund H (1995) Kinetics for the Life Sciences. Cambridge University Press, New YorkCrossRefGoogle Scholar
  10. Hartwig JH, Kwiatkowski DJ (1991) Actin-binding proteins. Curr Opin Cell Biol 3:87–97PubMedCrossRefGoogle Scholar
  11. Hiromi K (1979) Kinetics of fast reactions. John Wiley, New YorkGoogle Scholar
  12. Isenberg G (1995) Cytoskeleton proteins:a purification manual. Springer, Berlin Heidelberg New YorkGoogle Scholar
  13. Johnson RP, Craig SW (1995) F-actin binding site masked by the intramolecular association of vinculin head and tail domains. Nature 371:261–264CrossRefGoogle Scholar
  14. Kouyama T, Mihashi K (1981) Fluorimetry study of N-(1-pyrenyl)-iodoacetamide labelled F-actin. Eur J Biochem 114:33–38PubMedCrossRefGoogle Scholar
  15. Maciver SK (1995) Microfilament organization and actin binding proteins. In:Hesketh JE, Pryme I (eds) The cytoskeleton, vol 1. Structure and assembly. JAI Press, Greenwich, ConnecticutGoogle Scholar
  16. Pardee JD, Spudich JA (1982) Purification of muscle actin. In:Frederiksen DW, Cunningham LW (eds) Methods in Enzymology, vol 85. Academic Press, New York, pp 164–181Google Scholar

General Reading

  1. Kurstin K (ed) (1969) Methods in enzymology, vol XVI. Fast reactions. Academic Press, New YorkGoogle Scholar
  2. Nordlie RC (1982) Kinetic examination of enzyme mechanisms involving branched reaction pathways. In:Purich DL (ed) Methods in enzymology, vol 87. Academic Press, New York, pp 319–548Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • Wolfgang H. Goldmann
    • 1
  • Zeno Guttenberg
    • 1
  • Robert M. Ezzell
    • 1
  • Gerhard Isenberg
    • 2
  1. 1.Surgery Research Laboratories, Massachusetts General Hospital, Dept. of SurgeryHarvard Medical SchoolCharlestownUSA
  2. 2.Department of Biophysics E22Technical University of MunichGarchingGermany

Personalised recommendations