Investigation of Ultrasound Influence on Myoglobin Deposition from Solutions on Sers Substrates

The article deals with the problem of recording the Raman spectra of protein solutions with a concentration of less than 1 μg /ml, which appears very important at present. To increase the concentration and amplification of the Raman signal from these proteins, sample preparation was carried out by drying a drop of solution on the surface of the substrate, which amplifies the signal for the Raman proteins scattering. The ultrasound effect on the process of protein deposition on the surface and its conformation was studied. Moreover, we conducted experiments on the preparation of a protein solution of myoglobin in water at concentration of 1 μg /ml on substrates that are thin (not more than 100 nm) silver films with a nanoscale surface structure made on mica substrates. After ultrasonic treatment, crystal-like protein structures were obtained, the amplitude of the Raman spectra being much larger than the that of similar spectra obtained after drying solutions of protein of the same concentration under normal conditions without ultrasound. Findings of the research show that ultrasound affects the nature of aggregation, crystallization and conformation of proteins, the amplitude and shape of the Raman spectra

This work was supported by the grant of the RNF No. 16-14-00209 of January 11, 2016

References

[1] Crespo R., Martins P.M., Gales L., Rochac F., Damasa A.M. Potential use of ultrasound to promote protein crystallization. Journal of Applied Crystallography, 2010, vol. 43, no. 6, pp. 1419–1425. DOI: 10.1107/S0021889810040951 Available at: https://onlinelibrary.wiley.com/doi/full/10.1107/S0021889810040951

[2] McCausland L.J., Cains P.W. Power ultrasound — a means to promote and control crystallization in biotechnology. Biotechnology and Genetic Engineering Reviews, 2004, vol. 21, no. 1, pp. 3–10. DOI: 10.1080/02648725.2004.10648048 Available at: https://www.tandfonline.com/doi/abs/10.1080/02648725.2004.10648048

[3] Chirkina A.A. Modern problems of biochemistry. Methods of research. Art. 34 2013.

[4] Bennig G.K. Atomic force microscope and method for imaging surfaces with atomic resolution. Patent US 4724318. App. 09.02.1986, publ. 04.08.1988.

[5] Martin Y., Williams C.C., Wickramasinghe H.K. Method for imaging sidewalls by atomic force microscopy. Appl. Phys. Lett., 1994, vol. 64, no. 19, art. 2498. DOI: 10.1063/1.111578 Available at: https://aip.scitation.org/doi/abs/10.1063/1.111578

[6] Meyer G., Amer N.M. Novel optical approach to atomic force microscopy. Appl. Phys. Lett., 1988, vol. 53, no. 12, art. 1045. DOI: 10.1063/1.100061 Available at: https://aip.scitation.org/doi/abs/10.1063/1.100061

[7] Durig U., Gimzewski J.K., Pohl D.W. Experimental observation of forces acting during scanning tunneling microscopy. Phys. Rev. Lett., 1986, vol. 57, no. 19, art. 2403. DOI: 10.1103/PhysRevLett.57.2403 Available at: https://journals.aps.org/prl/abstract/10.1103/PhysRevLett.57.2403