Invited Speaker 23rd International Society of Magnetic Resonance Conference 2023

Gd(III)-19F distance measurements in solution and in cells by electron-nuclear double resonance (#30)

Daniellla Goldfarb 1
  1. Chemical and Biological Physics, Weizmann Institute of Sciene, Rehovit, Israel

Double electron-electron resonance (DEER)  can track proteins’ conformations by providing distance distributions between two attached spin labels. This technique, however, cannot access distances below 1.5 nm. Very recently, nitroxide-19F,[1] trityl-19F,[2] and Gd(III)-19F[3] electron-nuclear double resonance (ENDOR) have been shown to efficiently determine  distances below 1.5 nm. Here we present Gd(III)-19F Mims ENDOR distance measurements in the range of 0.9 nm to 1.5 nm on several model complexes and 19F labeled  GB1 and ubiquitin (Ub) proteins, spin-labeled with rigid Gd(III) tags. We further extended these studies to Gb1 and ubiquitin delivered into Hela cells, thus adding 19F ENDOR to the tool-box allowing exporlating protein in their native environment, the cell.4 The solution and in-cell derived Gd(III)-19F distances were essentially identical and lie in the 1-1.5 nm range revealing that both, GB1 and Ub, retained their overall structure in the cell. Methods for incresing the sensetivity of  19F Mims ENDOR and incresing the accessible distance range will be presented.

 

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