The Quiet Renaissance of Protein Nuclear Magnetic Resonance

Abstract

From roughly 1985 through the start of the new millennium, the cutting edge of solution protein nuclear magnetic resonance (NMR) spectroscopy was to a significant extent driven by the aspiration to determine structures. Here we survey recent advances in protein NMR that herald a renaissance in which a number of its most important applications reflect the broad problem-solving capability displayed by this method during its classical era during the 1970s and early 80s. “Without receivers fitted and kept in order, the air may tingle and thrill with the message, but it will not reach my spirit and consciousness.” Mary Slessor, Calabar, circa 1910

Engineered solubility tag for solution NMR of proteins

“…The approach presented here for increasing protein solubility has a number of obvious advantages: (1) there are a number of commercially available linkers that can be used that vary in chemical composition and length, 41 (2) the solubility tag can be introduced at any position on the target protein, (3) nearly all the target protein was labeled in the conjugation reaction, (4) ability to selectively introduce isotopic labels into the target protein, as well as deuterate the solubility tag, (5) the labeling reactions can be conducted under a wide range of solution conditions that are compatible with downstream NMR applications, and (6) highly sensitive fluorescence-based analysis determining whether or not the native fold is preserved upon labeling. The drawback of the method is the requirement for a solvent-exposed cysteine.…”

“…Solution nuclear magnetic resonance (NMR) has proven to be a powerful technique for studies of structure, stability, and dynamics of proteins . Over the last decade we have observed significant advances in hardware and methodologies that improve the sensitivity of solution NMR .…”

“…Solution nuclear magnetic resonance (NMR) has proven to be a powerful technique for studies of structure, stability, and dynamics of proteins. 1 Over the last decade we have observed significant advances in hardware and methodologies that improve the sensitivity of solution NMR. 2,3 While nowadays, 1D measurements and 2D 1 H- 15 N and 1 H- 13 C correlation maps can often be achieved at concentrations of tens of micromolar, a number of other experiments, such as backbone and side-chain assignments, studies of dynamics and acquisition of NOESY datasets for structure determination, require protein concentrations in excess of hundreds of micromolar.…”

Molecular basis of MAP kinase regulation

“…While X‐ray crystallography has provided essential insights into the regulation of MAPKs by their interacting proteins, over the last ∼10 years biomolecular NMR spectroscopy has also been used to complement and critically expand these efforts (Table ) . This contribution was accelerated by new protein labeling techniques, new, more sensitive NMR spectrometers as well as novel NMR techniques . To overcome the broad line‐widths that are characteristic of proteins ≥35 kDa, MAPKs must be expressed in D 2 O‐based medium and TROSY versions of all 2D and 3D experiments must be recorded using high field NMR spectrometers (800–1000 MHz 1 H Larmor frequency) .…”

Recent contributions of structure-based drug design to the development of antibacterial compounds

“…This fundamental property is harnessed in various NMR experiments to provide detailed information on the molecular structure and interactions of proteins [22]. Consequently, NMR is extensively used to assist antimicrobial drug development in various stages of the SBDD process including hit identification, hit validation, and lead optimization [23–25].…”