TY - JOUR
T1 - Structural design of intrinsically fluorescent oxysterols
AU - Nåbo, Lina J.
AU - Modzel, Maciej
AU - Krishnan, Kathiresan
AU - Covey, Douglas F.
AU - Fujiwara, Hideji
AU - Ory, Daniel S.
AU - Szomek, Maria
AU - Khandelia, Himanshu
AU - Wüstner, Daniel
AU - Kongsted, Jacob
N1 - Funding Information:
Computational resources were provided by the DeIC National HPC Center at the University of Southern Denmark through an Abacus 2.0 grant. J. K. thanks the Danish Council for Independent Research and the Villum Foundation for financial support. D. S. O. and D. F. C. acknowledge NIH HL067773 for funding and D. F. C. further acknowledges support from the Taylor Institute for Innovative Psychiatric Research.
Publisher Copyright:
© 2017
PY - 2018/5
Y1 - 2018/5
N2 - Oxysterols are oxidized derivatives of cholesterol with many important biological functions. Trafficking of oxysterols in and between cells is not well studied, largely due to the lack of appropriate oxysterol analogs. Intrinsically fluorescent oxysterols present a new route towards direct observation of intracellular oxysterol trafficking by fluorescence microscopy. We characterize the fluorescence properties of the existing fluorescent 25-hydroxycholesterol analog 25-hydroxycholestatrienol, and propose a new probe with an extended conjugated system. The location of both probes inside a membrane is analyzed and compared with that of 25-hydroxycholesterol using molecular dynamics simulations. The analogs’ one- and two-photon absorption properties inside the membrane are evaluated using electronic structure calculations with polarizable embedding. Due to predicted keto–enol tautomerisation of the new oxysterol analog, we also evaluate the keto form. Both analogs are found to be good probe candidates for 25-hydroxycholesterol, provided that the new analog remains in the enol-form. Only the new analog with extended conjugated system shows significant two-photon absorption, which is strongly enhanced by the presence of the membrane.
AB - Oxysterols are oxidized derivatives of cholesterol with many important biological functions. Trafficking of oxysterols in and between cells is not well studied, largely due to the lack of appropriate oxysterol analogs. Intrinsically fluorescent oxysterols present a new route towards direct observation of intracellular oxysterol trafficking by fluorescence microscopy. We characterize the fluorescence properties of the existing fluorescent 25-hydroxycholesterol analog 25-hydroxycholestatrienol, and propose a new probe with an extended conjugated system. The location of both probes inside a membrane is analyzed and compared with that of 25-hydroxycholesterol using molecular dynamics simulations. The analogs’ one- and two-photon absorption properties inside the membrane are evaluated using electronic structure calculations with polarizable embedding. Due to predicted keto–enol tautomerisation of the new oxysterol analog, we also evaluate the keto form. Both analogs are found to be good probe candidates for 25-hydroxycholesterol, provided that the new analog remains in the enol-form. Only the new analog with extended conjugated system shows significant two-photon absorption, which is strongly enhanced by the presence of the membrane.
KW - Electronic structure calculation
KW - Fluorescence
KW - Imaging
KW - Intracellular transport
KW - Oxysterol
KW - Polarizable embedding
UR - http://www.scopus.com/inward/record.url?scp=85044265000&partnerID=8YFLogxK
U2 - 10.1016/j.chemphyslip.2017.12.005
DO - 10.1016/j.chemphyslip.2017.12.005
M3 - Article
C2 - 29287905
AN - SCOPUS:85044265000
SN - 0009-3084
VL - 212
SP - 26
EP - 34
JO - Chemistry and Physics of Lipids
JF - Chemistry and Physics of Lipids
ER -