David Neuhaus

This book is a comprehensive treatment of the nuclear Overhauser effect (NOE), an important branch of NMR spectroscopy concerned with structural and conformational problems. It is the only text in this area since 1971. Since that time there have been tremendous changes in the area, particularly with the growth of experiments such as NOE difference spectroscopy, NOESY, the heteronuclear NOE, with applications to an ever-increasing variety of structural and conformational problems in chemistry and biochemistry. The book gives a very clear and integrated account of all these developments. It covers theory, experimental practice, and applications in a unified manner including the influence of exchange and spin-spin-coupling as part of its underlying concepts, and applications to molecules as diverse as cyclobutanes and proteins. The book is of interest primarily to postgraduate organic and inorganic chemists and biochemists who use the NOE in their research, but it is also useful to those who wish to learn the background to the technique, for instance computational chemists , biophysicists, and physical chemists. Although it describes and explains the latest developments, the treatment is suitable for those with only a graduate-level background in chemistry and spectroscopy.

Poly(ADP-ribose)polymerase 1 (PARP-1) is a key eukaryotic stress sensor that responds in seconds to DNA single-strand breaks (SSBs), the most frequent genomic damage. A burst of poly(ADP-ribose) synthesis initiates DNA damage response, whereas PARP-1 inhibition kills BRCA-deficient tumor cells selectively, providing the first anti-cancer therapy based on synthetic lethality. However, the mechanism underlying PARP-1's function remained obscure; inherent dynamics of SSBs and PARP-1's multi-domain architecture hindered structural studies. Here we reveal the structural basis of SSB detection and how multi-domain folding underlies the allosteric switch that determines PARP-1's signaling response. Two flexibly linked N-terminal zinc fingers recognize the extreme deformability of SSBs and drive co-operative, stepwise self-assembly of remaining PARP-1 domains to control the activity of the C-terminal catalytic domain. Automodification in cis explains the subsequent release of monomeric PARP-1 from DNA, allowing repair and replication to proceed. Our results provide a molecular framework for understanding PARP inhibitor action and, more generally, allosteric control of dynamic, multi-domain proteins.

Novel strategies for elucidation and classification of amino acid 1H-NMR spin systems in proteins were developed exploiting recently introduced two-dimensional NMR techniques such as phase-sensitive double-quantum-filtered correlated spectroscopy, relayed coherence transfer spectroscopy, double quantum spectroscopy and nuclear Overhauser spectroscopy. Due to the improved resolution in phase-sensitive spectra, the fine structure of cross peaks could be exploited as a powerful source of information for establishing 1H-1H connectivities. Principles for the interpretation of multiplet structures of absorption mode cross peaks are discussed. With these methods the 1H spin systems of rabbit liver metallothionein-2 were elucidated and classified according to amino acid types. Despite the intrinsically difficult situation arising from the unusual amino acid composition of this protein, a more complete characterization of the 1H spin systems prior to the step of sequential resonance assignments was achieved with the presently introduced methodology than was possible in earlier studies of proteins of similar size.