Sammanfattning

Using the case of the catalytic domain of MMP-12 in complex with the known inhibitor CGS27023A, a recently assembled 3D N-15-edited/N-14,C-12-filtered ROESY experiment is used to monitor and distinguish protein amide protons in fast exchange with bulk water from amide protons close to water molecules with longer residence times, the latter possibly reflecting water molecules of structural or functional importance. The N-15-edited/N-14,C-12-filtered ROESY spectra were compared to the original N-15-edited/N-14,C-12-filtered NOESY and the conventional amide-water exchange experiment, CLEANEX. Three protein backbone amide protons experiencing direct dipolar cross relaxation with water in the N-15-edited/N-14,C-12-filtered ROESY spectrum were assigned. In an ensemble of six crystal structures, two conserved water molecules within 3 of the three amide protons were identified. These two water molecules are buried into cavities in the protein surface and thus sufficiently slowed down by the protein topology to account for the observed dipolar interaction. Structural analysis of an ensemble of six crystal structures ruled out any exchange-relayed contributions for the amide-water interactions of interest.

Nyckelord

Protein-water interactions; Metalloproteases; MMP-12; NMR; Hydration; X-filtering

Publicerad i

Journal of Biomolecular NMR
2012, Volym: 53, nummer: 2, sidor: 85-92
Utgivare: SPRINGER

UKÄ forskningsämne

Läkemedelskemi


Publikationens identifierare

DOI: https://doi.org/10.1007/s10858-012-9624-7

Permanent länk till denna sida (URI)

https://res.slu.se/id/publ/99463