NMR studies of an immunomodulatory benzodiazepine binding to its molecular target on the mitochondrial F1F0‐ATPase |
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Authors: | Andrew C. Stelzer Richard W. Frazee Chad Van Huis Joanne Cleary Anthony W. Opipari Jr. Gary D. Glick Hashim M. Al‐Hashimi |
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Affiliation: | 1. Department of Biophysics, University of Michigan, Ann Arbor, MI 48109;2. Department of Chemistry, University of Michigan, Ann Arbor, MI 48109;3. Lycera Corporation, 46701 North Commerce Center Drive, Plymouth, MI 48170;4. Department of Obstetrics and Gynecology, University of Michigan, Ann Arbor, MI 48109 |
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Abstract: | Bz‐423 is an inhibitor of the mitochondrial F1F0‐ATPase, with therapeutic properties in murine models of immune diseases. Here, we study the binding of a water‐soluble Bz‐423 analog (5‐(3‐(aminomethyl)phenyl)‐7‐chloro‐ 1‐methyl‐3‐(naphthalen‐2‐ylmethyl)‐1H‐benzo][e][1,4]diazepin‐2(3H)‐one); (1) to its target subunit on the enzyme, the oligomycin sensitivity conferring protein (OSCP), by NMR spectroscopy using chemical shift perturbation and cross‐relaxation experiments. Titration experiments with constructs representing residues 1–120 or 1–145 of the OSCP reveals that (a) 1 binds to a region of the protein, at the minimum, comprising residues M51, L56, K65, V66, K75, K77, and N92, and (b) binding of 1 induces conformational changes in the OSCP. Control experiments employing a variant of 1 in which a key binding element on the small molecule was deleted; it had no perturbational effect on the spectra of the OSCP, which indicates that the observed changes with 1 represent specific binding interactions. Collectively, these data suggest that 1 might inhibit the enzyme through an allosteric mechanism where binding results in conformational changes that perturb the OSCP‐F1 interface resulting in disrupted communication between the peripheral stalk and the F1‐domain of the enzyme. © 2009 Wiley Periodicals, Inc. Biopolymers 29: 85–92, 2010. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com |
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Keywords: | binding allosteric bioener |
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