Abstract
To assess the effects of adsorption on protein structure, ultraviolet optical absorption spectra of myoglobin (Mb) bound to polydimethylsiloxane (PDMS) were measured. A flow cell, which enabled adsorption under controlled hydrodynamic conditions, was used in conjunction with a conventional spectrophotometer to obtain the spectra. Adsorption to PDMS reduced significantly the absorbance in the Soret region of the Mb spectrum, whereas the spectrum in the region near 280 nm was essentially unaffected. This result showed that disruption of the native structure of Mb occurs following interaction with PDMS. Furthermore, the change in the absorption spectrum may indicate loss of heme from the heme pocket of the adsorbed protein. Mb structure was altered from its solution configuration within fifteen min of contact with the surface. Exchange of adsorbed Mb with Mb in solution had little or no effect on the absorption spectrum of the surface-confined protein, indicating that exchange occurs only between conformationally altered species or between native species.
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- Acampora G., Hermans J., Jr Reversible denaturation of sperm whale myoglobin. I. Dependence on temperature, pH, and composition. J Am Chem Soc. 1967 Mar 29;89(7):1543–1547. doi: 10.1021/ja00983a001. [DOI] [PubMed] [Google Scholar]
- BRESLOW E., GURD F. R. Reactivity of sperm whale metmyoglobin towards hydrogen ions and p-nitrophenyl acetate. J Biol Chem. 1962 Feb;237:371–381. [PubMed] [Google Scholar]
- Darst S. A., Robertson C. R., Berzofsky J. A. Adsorption of the protein antigen myoglobin affects the binding of conformation-specific monoclonal antibodies. Biophys J. 1988 Apr;53(4):533–539. doi: 10.1016/S0006-3495(88)83133-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eaton W. A., Hochstrasser R. M. Single-crystal spectra of ferrimyoglobin complexes in polarized light. J Chem Phys. 1968 Aug 1;49(3):985–995. doi: 10.1063/1.1670263. [DOI] [PubMed] [Google Scholar]
- Hanania G. I., Yeghiayan A., Cameron B. F. Absorption spectra of sperm-whale ferrimyoglobin. Biochem J. 1966 Jan;98(1):189–192. doi: 10.1042/bj0980189. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herskovits T. T., Gadegbeku B., Jaillet H. On the structural stability and solvent denaturation of proteins. I. Denaturation by the alcohols and glycols. J Biol Chem. 1970 May 25;245(10):2588–2598. [PubMed] [Google Scholar]
- Herskovits T. T., Jaillet H., Gadegbeku B. On the structural stability and solvent denaturation of proteins. II. Denaturation by the ureas. J Biol Chem. 1970 Sep 10;245(17):4544–4550. [PubMed] [Google Scholar]
- Hochstrasser R. M., Negus D. K. Picosecond fluorescence decay of tryptophans in myoglobin. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4399–4403. doi: 10.1073/pnas.81.14.4399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Privalov P. L., Tiktopulo E. I., Venyaminov SYu, Griko YuV, Makhatadze G. I., Khechinashvili N. N. Heat capacity and conformation of proteins in the denatured state. J Mol Biol. 1989 Feb 20;205(4):737–750. doi: 10.1016/0022-2836(89)90318-5. [DOI] [PubMed] [Google Scholar]
- Schechter A. N., Epstein C. J. Spectral studies on the denaturation of myoglobin. J Mol Biol. 1968 Aug 14;35(3):567–589. doi: 10.1016/s0022-2836(68)80015-4. [DOI] [PubMed] [Google Scholar]
- Shen L. L., Hermans J., Jr Kinetics of conformation change of sperm-whale myoglobin. I. Folding and unfolding of metmyoglobin following pH jump. Biochemistry. 1972 May 9;11(10):1836–1841. doi: 10.1021/bi00760a016. [DOI] [PubMed] [Google Scholar]