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Title
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Trypsin mutants for structure-based drug design: expression, refolding and crystallisation.
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Authors
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D.Rauh,
S.Reyda,
G.Klebe,
M.T.Stubbs.
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Ref.
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Biol Chem, 2002,
383,
1309-1314.
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PubMed id
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Abstract
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New techniques in drug discovery are essential for the fast and efficient
development of novel innovative drugs to deal with the challenges of the future.
Structure determinations of various members of serine proteinases have provided
a basis for computer-based drug design within this class of enzymes. In many
proteins of interest, however, this course is blocked through a lack of suitable
crystals. As a strategy for circumventing such problems, we have investigated
the use of surrogate proteins for studying protein-ligand interactions. To test
the feasibility of this approach, we have chosen bovine trypsin as a scaffold to
reconstruct the ligand binding site of factor Xa. The simple modular design of
trypsin, its readiness to crystallise and straightforward handling lends itself
to such drug design by proxy. The expression, folding, purification,
crystallographic and kinetic characterisation of bovine trypsin forms with
factor Xa phenotype are presented.
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