Two-Site Internally Cooperative Mechanism for Enzyme Kinetics in a Hydrogel Forming Recombinant Protein.

A recombinant protein, ATCTA, consisting of three domains, α-helix (A), thrombin cleavage site (T), and water-soluble coil (C), forms hydrogels via the self-association of its flanking α-helices into tetrameric bundles, which act as cross-links for the hydrogel network. In the presence of thrombin, the hydrogel degrades due to the thrombin cleavage sites. To better understand the proteolysis reaction in ATCTA, we performed a series of kinetic experiments on the proteins ATC, CTA, CTATC, and ATCTA.