COMPOUNDS AND METHODS OF MODULATING MITOCHONDRIAL BIOENERGETIC EFFICIENCY THROUGH AN INTERACTION WITH ATP SYNTHASE (COMPLEX V) AND ITS SUBUNITS
The present invention provides, in part, methods of identifying compounds that can bind to an ATP synthase complex, increase bioenergetic efficiency, decrease oxygen consumption or the rate thereof, increase oxygen utilization efficiency, increase cell survival or any combination thereof, and methods of using compounds and/or identified compounds to increase bioenergetic efficiency, increase oxygen utilization efficiency, decrease oxygen consumption, increase cell survival, or any combination thereof.
1 . A method of identifying a compound that increases oxygen utilization efficiency comprising
contacting a compound that binds a mitochondrial ATP synthase complex with a mitochondria or a sub-mitochondrial vesicle; and
measuring oxygen utilization efficiency,
wherein when the measured oxygen utilization efficiency in the mitochondria or a sub-mitochondrial vesicle increases in the presence of the compound that binds the mitochondrial ATP synthase complex indicates that the compound is a compound that increases oxygen utilization efficiency.
2 . The method of claim 1 , further comprising identifying a compound that binds to the mitochondrial ATP synthase complex comprising:
contacting a test compound with the mitochondrial ATP synthase complex;
and identifying the test compound as the compound that binds the mitochondrial ATP synthase complex.
3 . The method of claim 1 , wherein the mitochondria are isolated mitochondria or the mitochondrial ATP synthase is an isolated mitochondrial ATP synthase complex.
4 . (canceled)
5 . The method of claim 1 , wherein a cell comprises the mitochondria and/or the mitochondrial ATP synthase complex.
6 - 12 . (canceled)
13 . The method of claim 1 further comprising heterologously expressing and isolating the mitochondrial ATP synthase complex prior to contacting the compound with the mitochondrial ATP synthase complex.
14 . The method of claim 1 , further comprising measuring ATP synthesis in the presence of the compound, wherein when the measured ATP synthesis is at least maintained in the presence of the compound indicates that the compound increases oxygen utilization efficiency and at least maintains ATP synthesis.
15 . The method of claim 1 , wherein said mitochondrial ATP synthase complex comprises an F1 head, a b subunit, an OSCP subunit, or a combination thereof.
16 . The method of claim 1 , wherein said compound binds to an F1 head, a b subunit, an OSCP subunit, or a combination thereof.
17 . The method of claim 1 , wherein said measuring oxygen utilization comprises comparing the compound to a positive or a negative control.
18 . (canceled)
19 . The method of claim 2 , wherein said identifying the test compound as an identified test compound that binds the mitochondrial ATP synthase complex comprises determining whether said test compound can competitively inhibit the binding of dexpramipexole to the mitochondrial ATP synthase complex.
20 . The method of claim 17 , wherein said positive control comprises a detectable label.
21 - 24 . (canceled)
25 . The method of claim 1 , wherein measuring oxygen utilization efficiency comprises measuring inhibition of mitochondrial conductance.
26 - 29 . (canceled)
30 . The method of claim 1 , further comprising identifying a compound as a compound that treats a neurodegenerative disease comprising contacting a subject with a neurodegenerative disease with the compound identified as a compound that that increases oxygen utilization efficiency,
wherein a compound that improves the condition of the subject inhibits the progression of the neurodegenerative disease, ameliorates the neurodegenerative disease indicates the compound as a compound that treats a neurodegenerative disease.
31 . The method of claim 1 , further comprising identifying a compound as a compound that is a neuroprotectant comprising contacting a subject with a neurodegenerative disease with the compound identified as a compound that that increases oxygen utilization efficiency,
wherein a compound that improves the condition of the subject inhibits the progression of the neurodegenerative disease, ameliorates the neurodegenerative disease indicates the compound as a compound that treats a neurodegenerative disease.
32 . A method of identifying a compound that increases oxygen utilization efficiency and at least maintains ATP synthesis comprising:
contacting a mitochondria or a sub-mitochondrial vesicle with a compound that binds to a mitochondrial ATP synthase complex and measuring oxygen utilization efficiency and ATP synthesis,
wherein a compound that increases oxygen utilization efficiency and enhances or increases ATP synthesis identifies the compound as a compound that increases oxygen utilization efficiency and at least maintains ATP synthesis.
33 . The method of claim 32 , further comprising determining whether said compound can inhibit the binding of dexpramipexole to the mitochondrial ATP synthase complex, wherein a compound that inhibits the binding of dexpramipexole to the mitochondrial ATP synthase complex identifies the compound as a compound that binds to the mitochondrial synthase complex.
34 . The method of claim 32 , further comprising comparing oxygen utilization efficiency and ATP synthesis in the presence of the compound to oxygen utilization efficiency and ATP synthesis in the presence of dexpramipexole, wherein a compound that at least maintains oxygen utilization efficiency and at least maintains ATP synthesis as compared to dexpramipexole identifies the test compound as a compound that increases oxygen utilization efficiency and at least maintains ATP synthesis.
35 - 47 . (canceled)
48 . The method of claim 32 , comprising heterologously expressing and isolating the mitochondrial ATP synthase complex prior to contacting the compound with the mitochondrial ATP synthase complex.
49 - 53 . (canceled)
54 . The method of claim 32 , further comprising identifying a compound as a compound that treats a neurodegenerative disease comprising contacting a subject with a neurodegenerative disease with the compound identified as a compound that that at least increases oxygen utilization efficiency and at least maintains ATP synthesis,
wherein a compound that improves the condition of the subject, inhibits the progression of the neurodegenerative disease, or ameliorates the neurodegenerative disease indicates the compound as a compound that treats a neurodegenerative disease.
55 . A method of identifying a compound that treats a neurodegenerative disease comprising:
contacting a subject with a neurodegenerative disease with a test compound, wherein said test compound is a compound that increases oxygen utilization efficiency, a compound that at least maintains ATP synthesis, and/or a compound that binds to a mitochondrial ATP synthase complex,
wherein a compound that improves the condition of the subject, inhibits the progression of the neurodegenerative disease, or ameliorates the neurodegenerative disease indicates the compound as a compound that treats a neurodegenerative disease.
56 . The method of claim 55 , further comprising identifying a compound that increases oxygen utilization efficiency, a compound that at least maintains ATP synthesis, and/or a compound that binds to a mitochondrial ATP synthase complex.
57 . The method of claim 55 , wherein said test compound is a compound that increases oxygen utilization efficiency, at least maintains ATP synthesis, and binds to a mitochondrial ATP synthase complex.
58 - 68 . (canceled)