Protein Science, Vol 3, Issue 9 1602-1604, Copyright © 1994 by Cold Spring Harbor Laboratory Press

FOR THE RECORD

Identification of a molecular switch that selects between two crystal forms of bovine pancreatic trypsin inhibitor

W. H. GALLAGHER and K. M. CROKER
Department of Chemistry, University of Wisconsin-Eau Claire, Eau Claire, Wisconsin 54702-4004

Two crystal forms of bovine pancreatic trypsin inhibitor are produced between pH 8.39 and 10.13 when crystals are grown at room temperature from solutions of 1.5 M potassium phosphate. Lower pH values favor the form II crystals, whereas higher pH values favor the form III. The transition from one crystal form to the other occurs at pH 9.35. We examined the crystal lattice contacts in both crystal forms and identified an unusual interaction we believe explains these observations. Spanning the crystallographic 2-fold axis in form III crystals, the Lys 41 side-chain amino nitrogens from 2 symmetry-related molecules are only 2.72 A apart, implying they are hydrogen bonded to one another. In form II crystals, the Lys 41 side-chain amino group is protonated and forms a salt bridge with a solventderived phosphate group. For the Lys 41 side-chain amino groups to hydrogen bond in form III crystals, at least 1 member of the pair must be deprotonated. The transition that occurs at pH 9.35 marks the pK(a) for deprotonation. In solution, the pK(a) for the Lys 41 side chain is around 10.8. The pK(a) for one of the interacting Lys 41 side chains in form III crystals is therefore shifted downward by about 1.5 pH units. The energy for lowering the pK(a) value comes from the many additional intermolecular hydrogen bonds that are present in form III crystals: 19 compared to only 8 in form II crystals.
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