In vitro covalent binding of nafenopin-CoA to human liver proteins.
Endogenous fatty acyl-CoAs play an important role in the acylation of proteins. A number of xenobiotic carboxylic acids are able to mimic fatty acids, forming CoA conjugates and acting as substrates in pathways of lipid metabolism. In this study nafenopin, a substrate for human hepatic fatty acid-CoA ligases, was chosen as a model compound to study xenobiotic acylation of human liver proteins. (3)H-nafenopin (+/- unlabeled palmitate) or (14)C-palmitate (+/- unlabeled nafenopin) were incubated for up to 120 min at 37 degrees C with ATP, CoA, and homogenate protein (1 mg/ml) from four individual human livers. Nafenopin covalently bound to proteins was detectable in all human livers and increased with time. Nafenopin adduct formation was directly proportional to nafenopin-CoA formation (r = 0.985, p < 0.05). Attachment of nafenopin to proteins involved both thioester and amide linkages with 76 and 24% of adducts formed with proteins > 100 and 50-100 kDa, respectively. Protein acylation by palmitate was also demonstrated. Palmitate significantly inhibited nafenopin-CoA formation by 29% but had no effect on nafenopin-CoA-mediated protein acylation. In contrast, nafenopin significantly inhibited protein palmitoylation by palmitoyl-CoA. This is the first study to demonstrate a direct relationship between xenobiotic-CoA formation, acylation of human liver proteins, and inhibition of endogenous palmitoylation. The ability of xenobiotics to acylate tissue proteins may have important biological consequences including perturbation of endogenous regulation of protein localization and function.
Sallustio
B C
BC
Department of Clinical Pharmacology, The Queen Elizabeth Hospital, Woodville, South Australia, 5011, Australia.
Nunthasomboon
S
S
Drogemuller
C J
CJ
Knights
K M
KM
D000214:Acyl Coenzyme A / Q000737:chemistry / Q000378:metabolism*; D000215:Acylation; D000293:Adolescent; D000328:Adult; D000577:Amides / Q000737:chemistry; D003066:Coenzyme A Ligases / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009255:Nafenopin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D010168:Palmitates / Q000378:metabolism; D011506:Proteins / Q000737:chemistry; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D013438:Sulfhydryl Compounds / Q000737:chemistry; D015262:Xenobiotics / Q000737:chemistry; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
D000214:Acyl Coenzyme A; D000577:Amides; D010168:Palmitates; D011506:Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D013438:Sulfhydryl Compounds; D015262:Xenobiotics; D009255:Nafenopin; C051515:nafenopin-coenzyme A; D003066:Coenzyme A Ligases; C090799:FAA2 protein, S cerevisiae; D000098859:Long-Chain-Fatty-Acid-CoA Ligase