Hosamani, Basavaprabhu ; Kale, Raju R. ; Sharma, Hemlata ; Wachtel, Ellen ; Kesselman, Ellina ; Danino, Dganit ; Friedman, Noga ; Sheves, Mordechai ; Namboothiri, Irishi N.N. ; Patchornik, Guy (2016) Membrane protein crystallization in micelles conjugated by nucleoside base-pairing: A different concept Journal of Structural Biology, 195 (3). pp. 379-386. ISSN 1047-8477
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Official URL: http://doi.org/10.1016/j.jsb.2016.06.021
Related URL: http://dx.doi.org/10.1016/j.jsb.2016.06.021
Abstract
The dearth of high quality, three dimensional crystals of membrane proteins, suitable for X-ray diffraction analysis, constitutes a serious barrier to progress in structural biology. To address this challenge, we have developed a new crystallization medium that relies on the conjugation of surfactant micelles via base-pairing of complementary hydrophobic nucleosides. Base-pairs formed at the interface between micelles bring them into proximity with each other; and when the conjugated micelles contain a membrane protein, crystal nucleation centers can be stabilized, thereby promoting crystal growth. Accordingly, two hydrophobic nucleoside derivatives – deoxyguanosine (G) and deoxycytidine (C), each covalently bonded to a 10 carbon chain were synthesized and added to an aqueous solution containing octyl β-D-thioglucopyranoside micelles. These hydrophobic nucleosides induced the formation of oil-rich globules after 2 days incubation at 19 °C or after a few hours in the presence of ammonium sulfate; however, phase separation was inhibited by 100 mM GMP. The presence of the membrane protein bacteriorhodopsin in the conjugated – micellar dispersion resulted in the growth within the colorless globules of a variety of purple crystals, the color indicating a functional protein. On this basis, we suggest that conjugation of micelles via base-pair complementarity may provide significant assistance to the structural determination of integral membrane proteins.
Item Type: | Article |
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Source: | Copyright of this article belongs to Elsevier Science. |
ID Code: | 121332 |
Deposited On: | 14 Jul 2021 11:19 |
Last Modified: | 14 Jul 2021 11:19 |
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