Presence of mannose phosphate on the epidermal growth factor receptor in A-431 cells.
In this report, we demonstrate a novel post-translational modification of the epidermal growth factor (EGF) receptor. This modification involves the presence of phosphate, previously thought to exist only on amino acid residues in the EGF receptor, on oligosaccharides of the receptor. We have utilized several independent approaches to determine that mannose phosphate is present on the EGF receptor in A-431 cells. Following metabolic labeling with 32P, immunoisolation of the EGF receptor, and digestion with Pronase radioactivity was determined to be present on high mannose type oligosaccharides by concanavalin A chromatography. Also, after acid hydrolysis of in vivo 32P-labeled EGF receptor, radioactivity was detected that co-migrated with mannose 6-phosphate on two-dimensional thin layer electrophoresis. This radiolabeled material co-eluted with a mannose 6-phosphate standard from a high pressure liquid chromatography anion exchange column. Last, an acid hydrolysate of [3H]mannose-labeled EGF receptor contained two radiolabeled fractions, as analyzed by thin layer electrophoresis, and the radioactivity in one of these fractions was substantially reduced by alkaline phosphatase treatment prior to electrophoresis. These experiments indicate that the mature EGF receptor in A-431 cells contains mannose phosphate. This is a novel modification for membrane receptors and has only been reported previously for lysosomal enzymes and a few secreted proteins.
Todderud
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Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232.
Carpenter
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D000818:Animals; D002460:Cell Line; D066246:ErbB Receptors / Q000096:biosynthesis / Q000235:genetics*; D006031:Glycosylation; D006600:Hexosephosphates / Q000302:isolation & purification*; D008358:Mannose / Q000378:metabolism; D008360:Mannosephosphates / Q000302:isolation & purification*; D010710:Phosphates / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D011499:Protein Processing, Post-Translational; D014316:Tritium
D006600:Hexosephosphates; D008360:Mannosephosphates; D010710:Phosphates; D010761:Phosphorus Radioisotopes; D014316:Tritium; C027693:mannose-6-phosphate; D066246:ErbB Receptors; D008358:Mannose