[5] Avidin and streptavidin

https://doi.org/10.1016/0076-6879(90)84259-JGet rights and content

Publisher Summary

This chapter discusses the fundamental molecular properties of avidin and streptavidin. Carbohydrate-free avidin can be obtained by use of deglycosylating enzymes and it has been shown to be present in significant amounts in some commercial preparations from which it may be separated by use of lectin columns. Improvement on the original use of biotinyl cellulose came with the introduction of iminobiotinyl derivatives of Sepharose that utilized the pH dependence of the binding 24 to achieve efficient elution. In iminobiotin, the ureido group becomes a guanidinium group; the form in which this is uncharged is strongly bound. The gene for streptavidin has been cloned and sequenced with the ultimate objective of using it in general expression systems for detecting and isolating fusion proteins. The stability is greatly enhanced by biotin binding, because the total free energy of binding is about 330 kJ/mol of tetramer. The dissociation constant for biotin is so low that it can be estimated only from the ratio of the rate constants for binding and exchange. The binding is accompanied by a red shift of the tryptophan spectrum and by a decrease in fluorescence, either of which can be used as the basis for quantitative assays.

References (62)

  • GyorgyP.
  • MeslarH.W. et al.

    J. Biol. Chem.

    (1978)
  • ChaietL. et al.

    Arch. Biochem. Biophys.

    (1964)
  • Fraenkel-ConratH. et al.

    Arch. Biochem. Biophys.

    (1952)
  • WakilS.J. et al.

    Biochem. Biophys. Acta

    (1958)
  • GreenN.M.

    Adv. Protein Chem.

    (1975)
  • GreenN.M. et al.

    J. Biol. Chem.

    (1972)
  • WilchekM. et al.

    Anal. Biochem.

    (1988)
  • FinnF.M. et al.

    J. Biol. Chem.

    (1980)
  • UpdykeT.V. et al.

    J. Immunol. Methods

    (1984)
  • McCormickD.B.

    Anal. Biochem.

    (1965)
  • BayerE.A. et al.

    J. Biochem. Biophys. Methods

    (1986)
  • HenriksonK.P. et al.

    Anal. Biochem.

    (1979)
  • KohanskiR.A. et al.

    J. Biol. Chem.

    (1985)
  • DimrothP.

    FEBS Lett.

    (1982)
  • HarmonF.

    Anal. Biochem.

    (1980)
  • SundelinJ. et al.

    J. Biol. Chem.

    (1985)
  • EloH.A. et al.

    Comp. Biochem. Physiol.

    (1981)
  • HuberR. et al.

    J. Mol. Biol.

    (1987)
  • GarlickR.K. et al.

    J. Biol. Chem.

    (1988)
  • HeitzmannH. et al.
  • BayerE.A. et al.

    Methods Biochem. Anal.

    (1980)
  • PinnE. et al.

    Eur. J. Biochem.

    (1982)
  • BruchR.C. et al.

    Biochemistry

    (1982)
  • WeberP.C. et al.

    Science

    (1989)
  • HendricksonW.A. et al.
  • ArgaranaC.E. et al.

    Nucleic Acids Res.

    (1986)
  • HillerY. et al.

    Biochem. J.

    (1987)
  • HofmannK. et al.
  • ...
  • ...
  • Cited by (0)

    View full text