Identification and Characterization of a Selective Radioligand for ELOVL6

• Published in: Journal of biochemistry (Tokyo) Vol. 146; no. 3; pp. 429 - 437
• Main Authors: Shimamura, Ken, Takahashi, Hidekazu, Kitazawa, Hidefumi, Miyamoto, Yasuhisa, Nagumo, Akira, Tang, Cheng, Dean, Dennis, Nagase, Tsuyoshi, Sato, Nagaaki, Tokita, Shigeru
• Format: Journal Article
• Language: English
• Published: England Japanese Biochemical Society 01.09.2009; Oxford University Press
• Subjects: Acetyltransferases - antagonists & inhibitors; Acetyltransferases - genetics; Acetyltransferases - metabolism; Acyl Coenzyme A - metabolism; Animals; Cell Line; Drug Evaluation, Preclinical; elongases; ELOVL6; Enzyme Inhibitors - chemical synthesis; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - metabolism; Fatty Acids - analysis; Gene Expression; Hepatocytes - chemistry; Hepatocytes - enzymology; Humans; Indoles - chemical synthesis; Indoles - chemistry; Indoles - metabolism; inhibitor; Inhibitory Concentration 50; Isoenzymes; Kinetics; ligand binding; Ligands; Mice; Microsomes - enzymology; Molecular Structure; Oxadiazoles - chemical synthesis; Oxadiazoles - chemistry; Oxadiazoles - metabolism; Palmitic Acid - metabolism; Pichia - genetics; Pichia - metabolism; Polymerase Chain Reaction; Protein Binding; radioligand; Radioligand Assay; Recombinant Fusion Proteins - antagonists & inhibitors; Recombinant Fusion Proteins - metabolism; Regression Analysis; Spectrometry, Mass, Electrospray Ionization; elongases; radioligand; ELOVL6; inhibitor; ligand binding
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/jb/mvp088

ELOVL6, a member of the elongation of very long-chain fatty acids (ELOVL) family, has recently been identified as the rate-limiting enzyme for the elongation of palmitoyl-CoA. ELOVL6 deficient mice are protected from high-fat diet induced insulin resistance, suggesting that ELOVL6 might be a promising target for the treatment of metabolic disorders. Despite the increasing interest in Elovl6 as a therapeutic target, the lack of chemical tools for this enzyme has limited further elucidation of the biochemical and pharmacological properties of ELOVL6. We have identified Compound-A, a potent inhibitor for ELOVL6, by screening our company library and subsequently optimizing hit compounds. Compound-A potently inhibited human and mouse ELOVL6 and displayed >100-fold greater selectivity for ELOVL6 over other ELOVL family members. Consistent with its potent and selective inhibitory activity toward ELOVL6, [³H]Compound-A bound to ELOVL6 with high affinity while showing no specific binding to other ELOVL enzymes. The observation that [³H]Compound-A bound to ELOVL6 in a palmitoyl-CoA-dependent manner in the absence of malonyl-CoA and NADPH suggests that Compound-A might recognize an enzyme-substrate complex, e.g. an acyl-enzyme intermediate. Collectively, these observations demonstrate that Compound-A and its tritiated form are useful tools for biochemical and pharmacological characterization of ELOVL6.