Identification of a proacrosin precursor in the cell-free translation of boar testicular poly(A)(+)-mRNA.
A cell-free translation system was used to determine the molecular mass of the protein component of precursor(s) to boar proacrosin. Poly(A)(+)-mRNA was extracted from freshly excised boar testis into phenol/chloroform, precipitated in chilled (-20 degrees C) ethanol, then translated in a cell-free, reticulocyte lysate system with Tran 35S-label. Analysis of the resulting products by SDS-PAGE followed by autoradiography demonstrated multiple bands of translated proteins. Both Western blotting and immunoprecipitation with a specific polyclonal antibody to boar proacrosin yielded a single major band with a relative molecular weight of approximately 64,000. These results suggest that proacrosin (Mr = 53,000-55,000), which contains both protein and carbohydrate moieties, results from the cellular processing of a proacrosin precursor molecule.
Yi
L S
LS
Department of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, Missouri 63110.
Erbs
P A
PA
Willand
J L
JL
Polakoski
K L
KL
D000176:Acrosin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D002474:Cell-Free System; D004792:Enzyme Precursors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D008970:Molecular Weight; D011061:Poly A / Q000235:genetics / Q000378:metabolism; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013552:Swine; D013737:Testis / Q000378:metabolism
D004792:Enzyme Precursors; D012333:RNA, Messenger; D011061:Poly A; C062266:preproacrosin; D000176:Acrosin