Methyl TROSY spectroscopy to study large biomolecular complexes

Abstract

Solution state NMR spectroscopy is a powerful technique in structural biology that can provide unique information regarding the structure, dynamics, and interactions of biomolecular complexes. For a long time, its experimental range was limited to proteins of modest size. However, in the recent decades, the applicability of the method has been extended such that assemblies with molecular weights far over 100 kDa became amenable to detailed analyses. The breakthroughs that enabled these advances include the development of TROSYbased NMR techniques and procedures to produce samples that are labeled in specific methyl groups. Here, we discuss these novel approaches to the study of high molecular weight systems, explaining briefly the theoretical background behind the advancements and giving several recent practical examples. The major applications of methyl TROSY NMR spectroscopy are mentioned: studies of intermolecular interactions, protein dynamics, and complex biomolecular structures. With all this, we substantiate our notion that NMR spectroscopy will continue to be a highly valuable and relevant method for investigating large biomolecular complexes that is complementary to other structural techniques.