Title
Post-synthetic Spin-Labeling of RNA through Click Chemistry for PELDOR Measurements
Author
Mark Kerzhner, Dinar Abdullin, Jennifer Wie˛cek, Hideto Matsuoka, Gregor Hagelueken, Olav Schiemann, Michael Famulok
Year
2016
Journal
Chemistry A European Journal
Abstract
Site-directed spin labeling of RNA based on click chemistry is used in combination with pulsed electron-electron double resonance (PELDOR) to benchmark a nitroxide spin label, called here dU˛ . We compare this approach with another established method that employs the rigid spin label C¸ m for RNA labeling. By using CD spectroscopy, thermal denaturation measurements, CW-EPR as well as PELDOR we analyzed and compared the influence of dU˛ and C¸m on a self-complementary RNA duplex. Our results demonstrate that the conformational diversity of dU˛ is significantly reduced near the freezing temperature of a phosphate buffer, resulting in strongly orientation-selective PELDOR time traces of the dU˛ -labeled RNA duplex.
Instrument
J-810
Keywords
Circular dichroism, Secondary structure, Biochemistry