Real-time NMR monitoring of biological activities in complex physiological environments.
Biological reactions occur in a highly organized spatiotemporal context and with kinetics that are modulated by multiple environmental factors. To integrate these variables in our experimental investigations of 'native' biological activities, we require quantitative tools for time-resolved in situ analyses in physiologically relevant settings. Here, we outline the use of high-resolution NMR spectroscopy to directly observe biological reactions in complex environments and in real-time. Specifically, we discuss how real-time NMR (RT-NMR) methods have delineated insights into metabolic processes, post-translational protein modifications, activities of cellular GTPases and their regulators, as well as of protein folding events.
Campbell Family Cancer Research Institute, Ontario Cancer Institute, Toronto, Ontario, Canada.
Matthew J
MJ
Smith
Campbell Family Cancer Research Institute, Ontario Cancer Institute, Toronto, Ontario, Canada.
Christopher B
CB
Marshall
In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Berlin, Germany.
Francois-Xavier
FX
Theillet
In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Berlin, Germany.
Andres
A
Binolfi
In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Berlin, Germany. Electronic address: selenko@fmp-berlin.de.
Philipp
P
Selenko
Campbell Family Cancer Research Institute, Ontario Cancer Institute, Toronto, Ontario, Canada; Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada. Electronic address: mikura@uhnresearch.ca.
Mitsuhiko
M
Ikura
D000818:Animals; D055162:Biocatalysis; D004867:Equipment Design; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000295:instrumentation / Q000379:methods*; D011487:Protein Conformation; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*
D011506:Proteins; D020558:GTP Phosphohydrolases