Purification and partial characterization of a malignancy-associated glycoprotein.
The isolation from cancer patient serum of a glycoprotein (Cc) associated with the presence of a variety of malignancies was previously reported. Although preliminary chemical and physical data indicated that Cc was different from identified circulating glycoproteins, subsequent immunological studies suggested that it was closely related to alpha 1-acid glycoprotein. Further analysis revealed the presence of two components in some Cc preparations and prompted a re-examination of the isolation and characterization data. In the present study, Cc was purified by a modified protocol which involved the use of pleural fluid obtained from individuals with cancer, and an alpha 1-acid glycoprotein antibody column to remove contaminating alpha 1-acid glycoprotein. Typically, the material not retained by the antibody column gave a single band with Mr 53,000 when subjected to sodium dodecyl sulfate-polyacrylamide electrophoresis. Amino terminal analysis revealed that the protein contained a blocked amino terminus, and carbohydrate analysis indicated that complex, asparagine-linked saccharide units were present. The product could be distinguished from alpha 1-acid glycoprotein and other previously described circulating glycoproteins by several criteria, including molecular weight, isoelectric point, and amino acid and carbohydrate composition. One of three preparations isolated had an apparent Mr of 59,000. Carbohydrate analysis as well as deglycosylation studies showed that the change in molecular weight was due to increased glycosylation.
Taddei-Peters
W C
WC
Department of Biological Chemistry, M.S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.
Bhavanandan
V P
VP
Davidson
E A
EA
D000327:Adsorption; D000596:Amino Acids / Q000032:analysis; D014408:Biomarkers, Tumor / Q000032:analysis / Q000302:isolation & purification*; D001826:Body Fluids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis / Q000302:isolation & purification*; D006031:Glycosylation; D007457:Iodine Radioisotopes; D037102:Lectins / Q000378:metabolism; D010472:Perchlorates; D010994:Pleura
D000596:Amino Acids; D014408:Biomarkers, Tumor; D002241:Carbohydrates; D006023:Glycoproteins; D007457:Iodine Radioisotopes; D037102:Lectins; D010472:Perchlorates