| MCA-AFRATDHG-{Lys(DNP)}; N-terminal: MCA (7-methoxycoumarin-4-yl-acetyl) fluorophore; C-terminal: (Lys)-DNP (2,4 dinitrophenyl) quencher; Fluorescence is released when OMA1 recognizes and cleaves the OPA1 8-merpeptide (fluorescence reporter) presumably at the RA site.
The peptide MCA-AFRATDHG-{Lys(DNP)} is commonly referred to as the OPA1 FRET Peptide. This peptide consists of a fluorescent donor group (MCA, 7-methoxycoumarin-4-acetic acid) conjugated to the N-terminus, followed by a specific amino acid sequence, AFRATDHG, and a modified lysine residue with a dinitrophenyl (DNP) acceptor group.
Usage and Applications:
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Drug Studies Targeting Protease Activity: The OPA1 FRET Peptide is widely used as a substrate for studying the activity of proteases, particularly those involved in mitochondrial dynamics and apoptosis regulation, such as OPA1 (Optic Atrophy 1). By monitoring changes in fluorescence resonance energy transfer (FRET) between the donor and acceptor groups upon cleavage of the peptide by proteases, researchers can quantify protease activity in vitro and in cell-based assays.
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Assessment of Mitochondrial Health: Since OPA1 is a key regulator of mitochondrial fusion and fission processes, alterations in its activity or expression levels are associated with mitochondrial dysfunction and various diseases, including neurodegenerative disorders and cancer. The OPA1 FRET Peptide enables researchers to assess the impact of drug candidates or genetic manipulations on mitochondrial dynamics and function, providing insights into potential therapeutic strategies targeting mitochondrial dysfunction.
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High-Throughput Screening Assays: The OPA1 FRET Peptide can be adapted for use in high-throughput screening assays to identify small molecules or compounds that modulate protease activity or mitochondrial dynamics. These assays can facilitate the discovery of novel drug candidates for treating conditions associated with mitochondrial dysfunction or aberrant protease activity.
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Mechanistic Studies of Disease Pathways: By employing the OPA1 FRET Peptide in conjunction with complementary techniques such as live-cell imaging and biochemical assays, researchers can elucidate the molecular mechanisms underlying disease pathogenesis, particularly in the context of mitochondrial dysfunction-related disorders. This peptide serves as a valuable tool for dissecting the roles of specific proteases and mitochondrial proteins in disease progression and identifying potential therapeutic targets.
Conclusion: The OPA1 FRET Peptide, consisting of a fluorescent donor group and a modified lysine acceptor linked by a specific amino acid sequence, is a versatile tool for studying protease activity and mitochondrial dynamics in drug studies and biomedical research. Its applications span from mechanistic studies of disease pathways to high-throughput screening assays for drug discovery, offering valuable insights into the development of therapeutic interventions targeting mitochondrial dysfunction-related disorders. |
