Abstract
The activity of ribonuclease A (RNase A) during adsorption onto molecular smooth mica increases from 16% to 78% in a period of 24 h when compared to its activity in free solution at pH 5 and 20 +/- 0.5 degrees C. From electropotential plots, the tertiary structure of RNase A, the characteristics of the mica surface, and direct measurements of the intermolecular forces between two adsorbed enzyme layers, a molecular explanation is offered for the changing activity with time. Initially, the RNase A molecules lie flat-on the mica with their smallest axis perpendicular to and their active site facing the surface. As adsorption proceeds, the molecules slowly reorient until at long times they lie end-on with their largest axis perpendicular to the surface and their active site partially exposed to the free solution. A translational diffusion process is indicated for the phase transition and molecular reorientation of the RNase A molecules.
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- Bernstein F. C., Koetzle T. F., Williams G. J., Meyer E. F., Jr, Brice M. D., Rodgers J. R., Kennard O., Shimanouchi T., Tasumi M. The Protein Data Bank: a computer-based archival file for macromolecular structures. J Mol Biol. 1977 May 25;112(3):535–542. doi: 10.1016/s0022-2836(77)80200-3. [DOI] [PubMed] [Google Scholar]
- Gilson M. K., Honig B. H. Calculation of electrostatic potentials in an enzyme active site. Nature. 1987 Nov 5;330(6143):84–86. doi: 10.1038/330084a0. [DOI] [PubMed] [Google Scholar]
- Israelachvili J., Pashley R. The hydrophobic interaction is long range, decaying exponentially with distance. Nature. 1982 Nov 25;300(5890):341–342. doi: 10.1038/300341a0. [DOI] [PubMed] [Google Scholar]
- Marra J., Israelachvili J. Direct measurements of forces between phosphatidylcholine and phosphatidylethanolamine bilayers in aqueous electrolyte solutions. Biochemistry. 1985 Aug 13;24(17):4608–4618. doi: 10.1021/bi00338a020. [DOI] [PubMed] [Google Scholar]
- Norde W. Adsorption of proteins from solution at the solid-liquid interface. Adv Colloid Interface Sci. 1986 Sep;25(4):267–340. doi: 10.1016/0001-8686(86)80012-4. [DOI] [PubMed] [Google Scholar]
- Pashley R. M., McGuiggan P. M., Ninham B. W., Evans D. F. Attractive forces between uncharged hydrophobic surfaces: direct measurements in aqueous solution. Science. 1985 Sep 13;229(4718):1088–1089. doi: 10.1126/science.4035349. [DOI] [PubMed] [Google Scholar]