Incorporation of myristate and palmitate into the sheep reticulocyte transferrin receptor: evidence for identical sites of labeling.
The ability of sheep reticulocytes and plasma membranes isolated from them to incorporate fatty acids into the transferrin receptor has been examined using both [3H]palmitate and [3H]myristate. Both fatty acids, when incorporated into the transferrin receptor, can be released by treating the protein with 1 M hydroxylamine at pH 7.0. After treatment of the 3H-acylated receptor with borohydride, an 3H-labeled alcohol is released, suggesting that the receptor-bound fatty acid is in thioester linkage. With both [3H]myristate and [3H]palmitate, Cleveland maps from immunoprecipitates of the transferrin receptor labeled in intact cells and isolated membranes show that identical peptides are labeled. No evidence was obtained for qualitatively different labeling with the two fatty acids. In intact reticulocytes, incorporation of [3H]palmitate into the transferrin receptor is approximately 3.5 times greater than the incorporation of [3H]myristate from equivalent concentrations of the labeled fatty acids. However, in isolated reticulocyte plasma membranes, there is much less difference between palmitate and myristate incorporation (with ATP) or between their acyl-CoA derivatives. The reason for the discrepancy between cells and membranes is unknown but may be due to the presence in intact cells of more than one enzyme for activating the fatty acids. Acylation of the receptor in isolated plasma membranes is fourfold greater with the CoA derivatives than with the free fatty acids. The fatty acid activating enzyme(s) as well as the acyltransferase(s) appear to be membrane bound in reticulocytes.
D000215:Acylation; D000818:Animals; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D002918:Chymotrypsin / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D007163:Immunosorbent Techniques; D019814:Myristic Acid; D009227:Myristic Acids / Q000097:blood*; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000097:blood*; D014176:Protein Biosynthesis; D011990:Receptors, Transferrin / Q000378:metabolism*; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism; D012756:Sheep; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014316:Tritium
D006898:Hydroxylamines; D009227:Myristic Acids; D010169:Palmitic Acids; D011990:Receptors, Transferrin; D013438:Sulfhydryl Compounds; D019814:Myristic Acid; D014316:Tritium; D019811:Hydroxylamine; D019308:Palmitic Acid; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C028344:glutamyl endopeptidase; D003545:Cysteine