Analytical Approaches to Alcohol Dehydrogenase Structures

  • Madalina T. Gheorghe
  • Ingemar Lindh
  • William J. Griffiths
  • Jan Sjövall
  • Tomas Bergman
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 372)

Abstract

Sequence analysis of polypeptides by Edman degradation is dependent on a free N-terminal α-amino group for the reaction with phenylisothiocyanate. However, alcohol dehydrogenases, like many other proteins, contain an acetylated N-terminal residue which blocks degradation (cf. Tsunasawa and Hirano, 1993). The conventional approach for blocked proteins involves enzymatic or chemical cleavage and reverse-phase HPLC-sepa-ration of fragments, followed by internal sequence analysis. The drawbacks associated with this technique are high protein consumption, long handling times and the fact that the N-terminal fragment remains inaccessible to Edman degradation. In this paper, we have tested direct chemical deblocking and applied it to both a synthetic peptide corresponding to the N-terminal segment of horse liver alcohol dehydrogenase and to the intact protein.

Keywords

Zinc Vortex Nickel Cobalt Helium 

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Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Madalina T. Gheorghe
    • 1
  • Ingemar Lindh
    • 1
  • William J. Griffiths
    • 1
  • Jan Sjövall
    • 1
  • Tomas Bergman
    • 1
  1. 1.Department of Medical Biochemistry and BiophysicsKarolinska InstitutetStockholmSweden

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