Membrane protein crystallization in micelles conjugated by nucleoside base-pairing: A different concept
| Autor: | Dganit Danino, Mordechai Sheves, Hemlata Sharma, Irishi N. N. Namboothiri, Basavaprabhu Hosamani, Ellina Kesselman, Noga Friedman, Ellen Wachtel, Raju R. Kale, Guy Patchornik |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Oligonucleotide Synthesis Conjugated system Crystallography X-Ray Micelle Deoxycytidine Cryo-Tem law.invention 03 medical and health sciences Protein structure Pulmonary surfactant Structural Biology law Organic chemistry Crystallization Biology Integral membrane protein Base Pairing Purification Micelles Crystallography biology Chemistry Deoxyguanosine Bacteriorhodopsin Engineered-Micelles Combinatorial chemistry 030104 developmental biology Membrane protein Bacteriorhodopsins biology.protein |
| Zdroj: | Journal of structural biology. 195(3) |
| ISSN: | 1095-8657 |
| Popis: | 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 beta-D-thioglucopyranoside micelles. These hydrophobic nucleosides induced the formation of oil-rich globules after 2 days incubation at 19 degrees 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. (C) 2016 Elsevier Inc. All rights reserved. |
| Databáze: | OpenAIRE |
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