Fast Mapping of Biomolecular Interfaces by Random Spin Labeling (RSL)

Maximilian J. Hartl Lehrstuhl Biopolymere, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany; Research Center for Bio-Macromolecules, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany

Björn M. Burmann Lehrstuhl Biopolymere, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany

Stefan J. Prasch Lehrstuhl Biopolymere, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany

Clemens Schwarzinger Institut für Chemische Technologie Organischer Stoffe, Johannes Kepler Universität, Altenbergerstraße 69, 4040, Linz, Austria

Kristian Schweimer Lehrstuhl Biopolymere, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany; Research Center for Bio-Macromolecules, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany

Birgitta M. Wöhrl Lehrstuhl Biopolymere, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany; Research Center for Bio-Macromolecules, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany

Paul Rösch Lehrstuhl Biopolymere, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany; Research Center for Bio-Macromolecules, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany

Stephan Schwarzinger Lehrstuhl Biopolymere, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany; Research Center for Bio-Macromolecules, Universität Bayreuth, Universitätsstraße 30, 95440, Bayreuth, Germany

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Abstract

Random spin labeling (RSL) is a method for rapid mapping of biomolecular interaction surfaces using an interaction partner with SL and an interaction partner enriched in ¹³C or ¹⁵N nuclei for paramagnetic relaxation enhanced NMR-based detection. The SL reaction is conducted in a manner resulting in a heterogeneous reaction product consisting of different populations of the protein carrying a varying number of spin labels at different positions. Preparation of the paramagnetic probe is complete within a few hours and hence much faster than site selective SL. RSL is applicable to tightly interacting systems but shows its particular strength when applied to systems involving weak or transient contacts.

Fast Mapping of Biomolecular Interfaces by Random Spin Labeling (RSL)

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Fast Mapping of Biomolecular Interfaces by Random Spin Labeling (RSL)

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