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Proc Natl Acad Sci U S A. Feb 1, 1992; 89(3): 1100–1104.

Enzymatic catalysis and dynamics in low-water environments.


Enzymes suspended in organic solvents represent a versatile system for studying the involvement of water in enzyme structure and function. Addition of less than 1% (vol/vol) water to tetrahydrofuran containing 1 M 1-propanol leads to a substantial increase in the transesterification activity of subtilisin Carlsberg (from Bacillus licheniformis) that correlates with a sharp increase in the active-site polarity and a 90% decrease in the rotational correlation time (i.e., increase in mobility) of a nitroxide spin label within the active site. Water in excess of 1% has little additional effect on active-site polarity and coincides with a further increase in spin-label mobility, yet the transesterification activity decreases dramatically. Thus, transesterification activity increases and then decreases with increasing enzyme hydration and flexibility (which are presumably coupled through dielectric screening), suggesting that the conformation of partially hydrated subtilisin is different from that of the nearly dry enzyme--i.e., enzyme containing less than 9% (wt/wt) water.

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Selected References

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  • Bone S, Pethig R. Dielectric studies of the binding of water to lysozyme. J Mol Biol. 1982 May 25;157(3):571–575. [PubMed]
  • Careri G, Gratton E, Yang PH, Rupley JA. Correlation of IR spectroscopic, heat capacity, diamagnetic susceptibility and enzymatic measurements on lysozyme powder. Nature. 1980 Apr 10;284(5756):572–573. [PubMed]
  • Careri G, Giansanti A, Rupley JA. Proton percolation on hydrated lysozyme powders. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6810–6814. [PMC free article] [PubMed]
  • Baker LJ, Hansen AM, Rao PB, Bryan WP. Effects of the presence of water on lysozyme conformation. Biopolymers. 1983 Jul;22(7):1637–1640. [PubMed]
  • Schinkel JE, Downer NW, Rupley JA. Hydrogen exchange of lysozyme powders. Hydration dependence of internal motions. Biochemistry. 1985 Jan 15;24(2):352–366. [PubMed]
  • Careri G, Giansanti A, Gratton E. Lysozyme film hydration events: an ir and gravimetric study. Biopolymers. 1979 May;18(5):1187–1203. [PubMed]
  • Bone S. Time-domain reflectrometry studies of water binding and structural flexibility in chymotrypsin. Biochim Biophys Acta. 1987 Nov 5;916(1):128–134. [PubMed]
  • Zaks A, Klibanov AM. Enzymatic catalysis in nonaqueous solvents. J Biol Chem. 1988 Mar 5;263(7):3194–3201. [PubMed]
  • Bott R, Ultsch M, Kossiakoff A, Graycar T, Katz B, Power S. The three-dimensional structure of Bacillus amyloliquefaciens subtilisin at 1.8 A and an analysis of the structural consequences of peroxide inactivation. J Biol Chem. 1988 Jun 5;263(16):7895–7906. [PubMed]
  • Morrisett JD, Broomfield CA. A comparative study of spin-labeled serine enzymes: acetylcholinesterase, trypsin, -chymotrypsin, elastase, and subtilisin. J Biol Chem. 1972 Nov 25;247(22):7224–7231. [PubMed]

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