Difference between revisions of "Cumero 2012 Br J Pharmacol"
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{{Publication | {{Publication | ||
|title=Cumero S, Fogolari F, Domenis R, Zucchi R, Mavelli I, Contessi S (2012) F(0) F(1) -Atpsynthase as a molecular target of 3-Iodothyronamine. Br J Pharmacol | |title=Cumero S, Fogolari F, Domenis R, Zucchi R, Mavelli I, Contessi S (2012) F(0) F(1) -Atpsynthase as a molecular target of 3-Iodothyronamine. Br J Pharmacol 166: 2331-2347 | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed?term=F0F1-Atpsynthase%20as%20a%20Molecular%20Target%20of%203-Iodothyronamine PMID: 22452346] | |info=[http://www.ncbi.nlm.nih.gov/pubmed?term=F0F1-Atpsynthase%20as%20a%20Molecular%20Target%20of%203-Iodothyronamine PMID: 22452346] | ||
|authors=Cumero S, Fogolari F, Domenis R, Zucchi R, Mavelli I, Contessi S | |authors=Cumero S, Fogolari F, Domenis R, Zucchi R, Mavelli I, Contessi S | ||
| Line 6: | Line 6: | ||
|journal=Br J Pharmacol | |journal=Br J Pharmacol | ||
|abstract=''Background & Purpose'': T1AM is a thyronamine derivative of thyroid hormone acting as a signalling molecule via non-genomic effectors and can reach intracellular targets. In light of the importance of F(0) F(1) -ATPsynthase as a target in drug development, T1AM interaction with the enzyme is demonstrated by its effects on the activity and a model of binding locations is depicted. | |abstract=''Background & Purpose'': T1AM is a thyronamine derivative of thyroid hormone acting as a signalling molecule via non-genomic effectors and can reach intracellular targets. In light of the importance of F(0) F(1) -ATPsynthase as a target in drug development, T1AM interaction with the enzyme is demonstrated by its effects on the activity and a model of binding locations is depicted. | ||
''Experimental Approach'': Kinetic analyses were performed on F(0) F(1) -ATPsynthase in sub-mitochondrial particles and soluble F(1) -ATPase. Activity assays and immunodetection of the inhibitor protein IF(1) were used and combined with molecular docking analyses. In situ respirometric analysis of T1AM effect was investigated on H9c2 cardiomyocytes. | ''Experimental Approach'': Kinetic analyses were performed on F(0) F(1) -ATPsynthase in sub-mitochondrial particles and soluble F(1) -ATPase. Activity assays and immunodetection of the inhibitor protein IF(1) were used and combined with molecular docking analyses. ''In situ'' respirometric analysis of T1AM effect was investigated on H9c2 cardiomyocytes. | ||
''Key Results'': T1AM is a non-competitive inhibitor of F(0) F(1) -ATPsynthase whose binding is mutually exclusive with that of the inhibitors IF(1) and aurovertin B. Distinct T1AM binding sites are consistent with results from both kinetic and docking analyses: at low nanomolar concentrations, T1AM binds to a high affinity-region likely located within the IF(1) binding site, causing IF(1) release; at higher concentrations, T1AM binds to a low affinity-region likely located within the aurovertin binding cavity and inhibits enzyme activity. Low nanomolar concentrations of T1AM elicit in cardiomyocytes an increase in ADP-stimulated mitochondrial respiration indicative for an activation of F(0) F(1) -ATPsynthase consistent with displacement of endogenous IF(1, ) thereby reinforcing the in vitro results. Conclusions & Implications: The T1AM effects upon F(0) F(1) -ATPsynthase are twofold: IF(1) displacement and enzyme inhibition. By targeting F(0) F(1) -ATPsynthase within mitochondria T1AM might affect cell bioenergetics with a positive effect on mitochondrial energy production at low endogenous concentration. T1AM putative binding locations overlapping with IF(1) and aurovertin binding sites are depicted. | ''Key Results'': T1AM is a non-competitive inhibitor of F(0) F(1) -ATPsynthase whose binding is mutually exclusive with that of the inhibitors IF(1) and aurovertin B. Distinct T1AM binding sites are consistent with results from both kinetic and docking analyses: at low nanomolar concentrations, T1AM binds to a high affinity-region likely located within the IF(1) binding site, causing IF(1) release; at higher concentrations, T1AM binds to a low affinity-region likely located within the aurovertin binding cavity and inhibits enzyme activity. Low nanomolar concentrations of T1AM elicit in cardiomyocytes an increase in ADP-stimulated mitochondrial respiration indicative for an activation of F(0) F(1) -ATPsynthase consistent with displacement of endogenous IF(1, ) thereby reinforcing the ''in vitro'' results. | ||
|keywords=H9c2 cardiomyocytes, | ''Conclusions & Implications'': The T1AM effects upon F(0) F(1) -ATPsynthase are twofold: IF(1) displacement and enzyme inhibition. By targeting F(0) F(1) -ATPsynthase within mitochondria T1AM might affect cell bioenergetics with a positive effect on mitochondrial energy production at low endogenous concentration. T1AM putative binding locations overlapping with IF(1) and aurovertin binding sites are depicted. | ||
|keywords=H9c2 cardiomyocytes, inhibitor of ATPsynthase, mitochondrial energy production | |||
|mipnetlab=IT Udine Grassi B, | |mipnetlab=IT Udine Grassi B, | ||
}} | }} | ||
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|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
|tissues=Cardiac muscle | |tissues=Cardiac muscle | ||
| | |couplingstates=OXPHOS, ETS | ||
|enzymes=Complex V; ATP Synthase | |||
|kinetics=Inhibitor; Uncoupler | |||
|topics=ATP; ADP; AMP; PCr | |||
}} | }} | ||
Revision as of 19:50, 26 July 2012
| Cumero S, Fogolari F, Domenis R, Zucchi R, Mavelli I, Contessi S (2012) F(0) F(1) -Atpsynthase as a molecular target of 3-Iodothyronamine. Br J Pharmacol 166: 2331-2347 |
Cumero S, Fogolari F, Domenis R, Zucchi R, Mavelli I, Contessi S (2012) Br J Pharmacol
Abstract: Background & Purpose: T1AM is a thyronamine derivative of thyroid hormone acting as a signalling molecule via non-genomic effectors and can reach intracellular targets. In light of the importance of F(0) F(1) -ATPsynthase as a target in drug development, T1AM interaction with the enzyme is demonstrated by its effects on the activity and a model of binding locations is depicted. Experimental Approach: Kinetic analyses were performed on F(0) F(1) -ATPsynthase in sub-mitochondrial particles and soluble F(1) -ATPase. Activity assays and immunodetection of the inhibitor protein IF(1) were used and combined with molecular docking analyses. In situ respirometric analysis of T1AM effect was investigated on H9c2 cardiomyocytes. Key Results: T1AM is a non-competitive inhibitor of F(0) F(1) -ATPsynthase whose binding is mutually exclusive with that of the inhibitors IF(1) and aurovertin B. Distinct T1AM binding sites are consistent with results from both kinetic and docking analyses: at low nanomolar concentrations, T1AM binds to a high affinity-region likely located within the IF(1) binding site, causing IF(1) release; at higher concentrations, T1AM binds to a low affinity-region likely located within the aurovertin binding cavity and inhibits enzyme activity. Low nanomolar concentrations of T1AM elicit in cardiomyocytes an increase in ADP-stimulated mitochondrial respiration indicative for an activation of F(0) F(1) -ATPsynthase consistent with displacement of endogenous IF(1, ) thereby reinforcing the in vitro results. Conclusions & Implications: The T1AM effects upon F(0) F(1) -ATPsynthase are twofold: IF(1) displacement and enzyme inhibition. By targeting F(0) F(1) -ATPsynthase within mitochondria T1AM might affect cell bioenergetics with a positive effect on mitochondrial energy production at low endogenous concentration. T1AM putative binding locations overlapping with IF(1) and aurovertin binding sites are depicted. • Keywords: H9c2 cardiomyocytes, inhibitor of ATPsynthase, mitochondrial energy production
• O2k-Network Lab: IT Udine Grassi B
Labels:
Tissue;cell: Cardiac muscle"Cardiac muscle" is not in the list (Heart, Skeletal muscle, Nervous system, Liver, Kidney, Lung;gill, Islet cell;pancreas;thymus, Endothelial;epithelial;mesothelial cell, Blood cells, Fat, ...) of allowed values for the "Tissue and cell" property.
Enzyme: Complex V; ATP Synthase"Complex V; ATP Synthase" is not in the list (Adenine nucleotide translocase, Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase, Inner mt-membrane transporter, Marker enzyme, Supercomplex, TCA cycle and matrix dehydrogenases, ...) of allowed values for the "Enzyme" property. Regulation: ATP; ADP; AMP; PCr"ATP; ADP; AMP; PCr" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property. Coupling state: OXPHOS, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property.
HRR: Oxygraph-2k
