Janovjak, Harald LIST Austria ; Kessler, Max; Oesterhelt, Dieter; Gaub, Hermann; Mueller, Daniel J
The combination of high-resolution atomic force microscopy (AFM) imaging and single-molecule force-spectroscopy was employed to unfold single bacteriorhodopsins (BR) from native purple membrane patches at various physiologically relevant temperatures. The unfolding spectra reveal detailed insight into the stability of individual structural elements of BR against mechanical unfolding. Intermittent states in the unfolding process are associated with the stepwise unfolding of alpha-helices, whereas other states are associated with the unfolding of polypeptide loops connecting the alpha-helices. It was found that the unfolding forces of the secondary structures considerably decreased upon increasing the temperature from 8 to 52°C. Associated with this effect, the probability of individual unfolding pathways of BR was significantly influenced by the temperature. At lower temperatures, transmembrane alpha-helices and extracellular polypeptide loops exhibited sufficient stability to individually establish potential barriers against unfolding, whereas they predominantly unfolded collectively at elevated temperatures. This suggests that increasing the temperature decreases the mechanical stability of secondary structural elements and changes molecular interactions between secondary structures, thereby forcing them to act as grouped structures.
5220 - 5229
Janovjak HL, Kessler M, Oesterhelt D, Gaub H, Mueller D. Unfolding pathways of native bacteriorhodopsin depend on temperature. EMBO Journal. 2003;22(19):5220-5229. doi:10.1093/emboj/cdg509
Janovjak, H. L., Kessler, M., Oesterhelt, D., Gaub, H., & Mueller, D. (2003). Unfolding pathways of native bacteriorhodopsin depend on temperature. EMBO Journal, 22(19), 5220–5229. https://doi.org/10.1093/emboj/cdg509
Janovjak, Harald L, Max Kessler, Dieter Oesterhelt, Hermann Gaub, and Daniel Mueller. “Unfolding Pathways of Native Bacteriorhodopsin Depend on Temperature.” EMBO Journal 22, no. 19 (2003): 5220–29. https://doi.org/10.1093/emboj/cdg509.
H. L. Janovjak, M. Kessler, D. Oesterhelt, H. Gaub, and D. Mueller, “Unfolding pathways of native bacteriorhodopsin depend on temperature,” EMBO Journal, vol. 22, no. 19, pp. 5220–5229, 2003.
Janovjak HL, Kessler M, Oesterhelt D, Gaub H, Mueller D. 2003. Unfolding pathways of native bacteriorhodopsin depend on temperature. EMBO Journal. 22(19), 5220–5229.
Janovjak, Harald L., et al. “Unfolding Pathways of Native Bacteriorhodopsin Depend on Temperature.” EMBO Journal, vol. 22, no. 19, Wiley-Blackwell, 2003, pp. 5220–29, doi:10.1093/emboj/cdg509.
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