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The high-resolution structures of a large number of adducts of matrix metalloproteinase 12 (MMP12) have helped in understanding the energetics of ligand-protein interactions. The analysis is applicable for any drug target for which structural information on adducts with a series of homologous ligands can be obtained, while the structural information obtained from 'in silico' docking is not very accurate.

The two missing copper ligands in the superoxide dismutase (SOD) like protein of Bacillus subtilis is reintroduced stepwise through site-directed mutagenesis. The catalytic properties of the double mutant, which might be regarded as a step in an artificial evolution from a nonactive SOD to a fully functioning enzyme, are discussed on the basis of the structural and dynamical properties.

Several studies are conducted to examine the reciprocal reorientation of the catalytic and hemopexin-like domains of full-length matrix metalloproteinases (MMP-12). MMP-1 and MMP-2 are shown to exhibit a conserved and well-defined spatial relationship between them.

In-cell NMR spectroscopy is a powerful approach to study protein structure and function in the native cellular environment. It provides precious insights into the folding, maturation, interactions, and ligand binding of important pharmacological targets directly in human cells. However, its widespread application is hampered by the fact that soluble globular proteins often interact with large cellular components, causing severe line broadening in conventional heteronuclear NMR experiments. 19 F NMR can overcome this issue, as fluorine atoms incorporated in proteins can be detected by simple background-free 1D NMR spectra. Here, we show that fluorinated amino acids can be easily incorporated in proteins expressed in human cells by employing a medium switch strategy. This straightforward approach allows the incorporation of different fluorinated amino acids in the protein of interest, reaching fluorination efficiencies up to 60%, as confirmed by mass spectrometry and X-ray crystallography. The versatility of the approach is shown by performing 19 F in-cell NMR on several proteins, including those that would otherwise be invisible by 1 H- 15 N in-cell NMR. We apply the approach to observe the interaction between an intracellular target, carbonic anhydrase 2, and its inhibitors, and to investigate how the formation of a complex between superoxide dismutase 1 and its chaperone CCS modulates the interaction of the chaperone subunit with the cellular environment.