Probing the conformation and orientation of adsorbed enzymes using side-chain modification.

ABSTRACT: The bioactivity of enzymes that are adsorbed on surfaces can be substantially influenced by the orientation of the enzyme on the surface and adsorption-induced changes in the enzyme's structure. Circular dichroism (CD) is a powerful method for observing the secondary structure of proteins; however, it provides little information regarding the tertiary structure of a protein or its adsorbed orientation. In this study, we developed methods using side-chain-specific chemical modification of solvent-exposed tryptophan residues to complement CD spectroscopy and bioactivity assays to provide greater detail regarding whether changes in enzyme bioactivity following adsorption are due to adsorbed orientation and/or adsorption-induced changes in the overall structure. These methods were then applied

SUBMITTER: Fears KP 

PROVIDER: S-EPMC3830457 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature