Biochemical Characterization of Theromin, a Novel Leech Inhibitor, and the Interaction between the Fourth Metalbinding Domain of Wilson Disease Protein and Its Copper(I) Chaperone HAH1
Date of Award
Doctor of Philosophy
Dr. David Huffman
Dr. Susan Stapleton
Dr. Ekkehard Sinn
Dr. Thomas Thamman
Theromin, a novel thrombin inhibitor from the leech Theromyzon tessulatum, has been shown to be the strongest inhibitor to date. There has been a lot of attention on direct thrombin inhibitors as a much more efficient drug for anticoagulation therapy. One main reason for the need of strong direct thrombin inhibitors is that the current therapy, heparin, causes alternative clotting events in some patients where antibodies are made to the heparin/thrombin complex and causes clots to occur by different methods. Recently there has been a strong inhibitor from a leech used, hirudin, that can inhibit with a Ki of 21 fM. This is the strongest inhibitor currently used as a therapy for patients suffering from clotting issues. This work investigated a leech inhibitor that has a Ki of 13 fM making it an even better candidate for therapies as it will inhibit more efficiently.
Copper is scrupulously regulated within the cell to maintain proper balance of redox activity and storage. One protein involved in this process is Wilson disease protein, which is responsible for moving copper into vesicles for either storage or removal from the cell. Previous work has shown copper is transferred to the Wilson disease protein via its metallochaperone, HAHl. The proposed mechanism for this transfer seems to involve a transient 3-coordinate copper(I) intermediate. Similar work done on Saccharomyces cerevisiae has shown that a 3-coordinate intermediate could exist between Atxl (analogue of HAHl) and its partner Ccc2. Trapping this intermediate involved selective mutation of individual cysteines involved in the copper(I) binding.
Zeider, Brian A., "Biochemical Characterization of Theromin, a Novel Leech Inhibitor, and the Interaction between the Fourth Metalbinding Domain of Wilson Disease Protein and Its Copper(I) Chaperone HAH1" (2010). Dissertations. 637.