ATP- and coenzyme A-dependent fatty acid incorporation into proteins of cell-free extracts from mouse tissues.
The incorporation of tritiated fatty acids into proteins has been studied in cell-free extracts from mouse tissues. Incubation of heart extracts with [3H]tetradecanoic or [3H]palmitic acid in the presence of ATP and CoA resulted in the time-dependent and selective labeling of proteins (Mr = 60,000, 47,000, 42,000, 31,000, 16,000, and 13,000) which could be detected after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Two polypeptides (Mr = 47,000 and 42,000) reached a maximum in fatty acid incorporation very rapidly and were mainly localized in the membrane subcellular fractions of the extract. These proteins underwent transient labeling with [3H] tetradecanoyl-CoA, the maximum incorporation being obtained within 1 min. The fatty acid-labeled proteins from tissue extracts had the same properties as other proteins known to be acylated in intact cells, i.e. the acyl moiety was resistant to delipidation with organic solvents but could be hydrolyzed by treatment with neutral hydroxylamine. Screening of different tissues showed that extracts from liver and kidney also catalyze the ATP- and CoA-dependent formation of a similar group of fatty acid-acylated proteins. The results provide evidence for a group of proteins in mammalian tissues which selectively incorporate fatty acids in vitro and should be of value for further studies on the biosynthesis of acylated proteins.
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002474:Cell-Free System; D003065:Coenzyme A / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005227:Fatty Acids / Q000378:metabolism*; D005470:Fluorometry; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D009206:Myocardium / Q000378:metabolism; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors
D005227:Fatty Acids; D006898:Hydroxylamines; D010169:Palmitic Acids; D011506:Proteins; D019811:Hydroxylamine; D019308:Palmitic Acid; D008623:Mercaptoethanol; D000255:Adenosine Triphosphate; D003065:Coenzyme A