TY - JOUR
T1 - Membrane protein crystallization in micelles conjugated by nucleoside base-pairing
T2 - A different concept
AU - Hosamani, Basavaprabhu
AU - Kale, Raju R.
AU - Sharma, Hemlata
AU - Wachtel, Ellen
AU - Kesselman, Ellina
AU - Danino, Dganit
AU - Friedman, Noga
AU - Sheves, Mordechai
AU - Namboothiri, Irishi N.N.
AU - Patchornik, Guy
N1 - Publisher Copyright:
© 2016 Elsevier Inc.
PY - 2016/9/1
Y1 - 2016/9/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84991387548&partnerID=8YFLogxK
U2 - 10.1016/j.jsb.2016.06.021
DO - 10.1016/j.jsb.2016.06.021
M3 - 文章
C2 - 27368128
AN - SCOPUS:84991387548
SN - 1047-8477
VL - 195
SP - 379
EP - 386
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 3
ER -