Purification, from cultured human choriocarcinoma cells, of a 75000-Mr protein reacting with antibodies to a synthetic peptide based on a cloned human endogenous provirus nucleotide sequence.
We previously detected in cultured choriocarcinoma cells a 75000-Mr polypeptide defined by immunoblotting with antibody to a synthetic peptide Sp23 (Cys-Glu-Asn-Pro-Ser-Gln-Phe-Tyr-Glu-Asp-Leu) based on a cloned human endogenous proviral nucleotide sequence. On immunohistological staining, anti-Sp23 stains antigen(s) in the syncytiotrophoblasts of first-trimester placentas and in renal-cell adenocarcinoma tissues. The present report describes purification to homogeneity of the protein from cultured choriocarcinoma cells. The procedure involves extraction with non-ionic detergent and h.p.l.c. using, sequentially, gel-permeation, anion-exchange and reverse-phase columns. The yield was 110 micrograms/g of total choriocarcinoma-cell protein. The results indicate that the purified protein is a monomeric and relatively hydrophilic molecule of Mr 75000.
D000906:Antibodies / Q000276:immunology*; D001483:Base Sequence; D002478:Cells, Cultured; D002822:Choriocarcinoma / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D004279:DNA, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D009363:Neoplasm Proteins / Q000276:immunology / Q000302:isolation & purification*; D009842:Oligopeptides / Q000276:immunology*; D011247:Pregnancy
D000906:Antibodies; D004279:DNA, Viral; D009363:Neoplasm Proteins; D009842:Oligopeptides; C046872:cysteinyl-glutamyl-asparaginyl-prolyl-serinyl-glutaminyl-phenylalanyl-tyrosyl-glutamyl-aspartyl-leucine