Peptide sequence analysis by nuclear magnetic resonance spectroscopy.

Abstract

Nuclear magnetic resonance spectroscopy (NMR) has found wide application to biochemical problems. This is reflected in the number of books (61, 130, 212, 243, 473) and review articles (95, 96, 163, 169, 294, 300, 325, 375) that describe this area of research. In recent years, considerable progress has been made in studies of the structure in solution of peptides, proteins, nucleotides and other biomolecules. Major technical developments were responsible for this success. These include: (1) the manufacture of superconducting magnets, (2) the general availability of Fourier transform NMR spectrometers, and (3) the discovery of paramagnetic chemical shift and relaxation enhancement (broadening) agents. In tandem, these advancements have extended the sensitivity and scope of NMR spectroscopy considerably. As a consequence, carbon-13 resonance studies no longer require an isotopic enrichment step (257, 420), and the available resolution has been improved to such a level that even the details of multiplet structure in protein 1H-spectra may be discerned (77–79, 130). The paramagnetic chemical shift and broadening probes may be used to simplify complex NMR spectra, and are useful in making peak assignments. Roberts and Jardetzky (375), in 1970, concluded that sequence determinations by NMR would likely be of limited value because of the inability to resolve adquately the resonances of 1H nuclei of the various amino acid residues. In this chapter, we will show that advances in NMR technology have generated renewed interest in peptide sequence analysis, and that this application of NMR has considerable merit and promise.