Identification of the primary translation product of atrial natriuretic peptide receptor mRNA in a cell-free system using anti-receptor antiserum.
The primary translation product of mRNA encoding atrial natriuretic peptide (ANP) receptor has been shown to have an Mr of 58,000. Poly(A)+ RNA was isolated from the bovine kidney and lung and translated in a rabbit reticulocyte lysate system containing [35S]methionine. Immunoprecipitation of the labeled translation products, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography, identified a 58-kDa protein as the primary translation product which is the unglycosylated precursor to be processed to the glycosylated mature 70-kDa form found in the plasma membranes. The result lends strong support to our previous proposal that mature ANP receptor is composed of two disulfide-linked 70-kDa subunits, eliminating the possibility that the two 70-kDa subunits arise from a larger 140-kDa precursor by proteolytic cleavage.
D000818:Animals; D002417:Cattle; D002474:Cell-Free System; D006023:Glycoproteins / Q000096:biosynthesis; D007106:Immune Sera; D008970:Molecular Weight; D011061:Poly A / Q000235:genetics*; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000235:genetics / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000235:genetics*; D017461:Receptors, Atrial Natriuretic Factor; D011956:Receptors, Cell Surface / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D006023:Glycoproteins; D007106:Immune Sera; D011498:Protein Precursors; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D011061:Poly A; D017461:Receptors, Atrial Natriuretic Factor