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| Joanne Yeh - | |
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Additives can improve the quality of macromolecular crystals
by presumably enhancing crystal contacts, conformational stability,
and/or solvent interactions. However, despite their widespread use,
the mechanisms by which these compounds exert their effects remain
largely unknown. We have used isothermal titration calorimetry and
differential scanning calorimetry to characterize the effects of
additives. The results of this study on five proteins found that the
positive effects of an additive are correlated with stabilization of
the macromolecule, as evidenced by shifts in the thermodynamic
variables, DH and DG of binding and on the midpoint melting transition
temperature, Tm. This approach of utilizing ultrasensitive
instrumental methods may allow rapid screening of a large number of
additives prior to crystallization and may be useful in assessing the
effect of a compound on the crystallizability of macromolecules.
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