EPR Relaxation‐Enhancement‐Based Distance Measurements on Orthogonally Spin‐Labeled T4‐Lysozyme
作者： Sahand Razzaghi , Evan K. Brooks , Enrica Bordignon , Wayne L. Hubbell , Maxim Yulikov
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摘要: Lanthanide-induced enhancement of the longitudinal relaxation nitroxide radicals in combination with orthogonal site-directed spin labeling is presented as a systematic distance measurement method intended for studies bio-macromolecules and bio-macromolecular complexes. The approach tested on water-soluble protein (T4-lysozyme) two different commercially available lanthanide labels, complemented by previously reported data membrane-inserted polypeptide. Single temperature measurements are shown to be sufficient reliable determination, an upper measurable limit about 5-6 nm. extracted averaged distances represent closest Ln(III) -nitroxide distributions. Studies conformational changes bio-macromolecule association-dissociation proposed possible application area relaxation-enhancement-based measurements.
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