Identification and partial characterization of soluble and membrane-bound KDN(deaminoneuraminic acid)-glycoproteins in human ovarian teratocarcinoma PA-1, and enhanced expression of free and bound KDN in cells cultured in mannose-rich media.
KDN (Deaminoneuraminic acid, or deaminated neuraminic acid) is a minor but biosynthetically independent member of the sialic acid. Human occurrence of KDN has already been established, although its level is so little that it is often undetectable by conventional sialic acid analysis. Elevated expression of KDN in fetal cord blood cells and some malignant tumor cells have been reported. However, in mammalian cells and tissues KDN mostly occurs as the free sugar and little occurred conjugated to glycolipids and/or glycoproteins. A positive correlation between the ratio of free KDN/free Neu5Ac in ovarian adenocarcinomas and the stage of malignancy has been noted for diagnostic use. We hypothesized that elevated expression of KDN in mammalian systems may be closely related to elevated activities of enzymes involved in the formation of sialoglycoconjugates and/or aberrant supply of the precursor sugar, mannose, used in the biosynthesis of KDN. In this study we used human ovarian teratocarcinoma cells PA-1 to further analyze KDN expression in human cells. Major findings reported in this paper are, (i) a 30 kDa KDN-glycoprotein immunostainable with monoclonal antibody, mAb.kdn3G, (specific for the KDNalpha2 --> 3Galbeta1--> epitope) and sensitive to KDNase was identified in the membrane fraction of the cell: (ii) a 49 kDa KDN-glycoprotein that is not reactive with mAb.kdn3G but is sensitive to KDNase was identified in the soluble fraction: and (iii) PA-1 cells showed unique response to mannose added to the growth medium in that the levels of both free and bound forms of KDN are elevated. This is the first report on the identification of mammalian KDN-glycoproteins by chemical and biochemical methods.
Inoue
Sadako
S
Institute of Biological Chemistry, Academia Sinica, Taipei 115-29, Taiwan.
Poongodi
Geetha L
GL
Suresh
Nimmagadda
N
Chang
Tschining
T
Inoue
Yasuo
Y
D001665:Binding Sites; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003470:Culture Media; D005260:Female; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D008358:Mannose / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000737:chemistry*; D012995:Solubility; D013400:Sugar Acids / Q000737:chemistry* / Q000378:metabolism; D018243:Teratocarcinoma / Q000737:chemistry*
D003470:Culture Media; D006023:Glycoproteins; D013400:Sugar Acids; C050008:3-deoxyglycero-galacto-nonulosonic acid; D008358:Mannose