Effect of desialylation of highly purified isoforms of human luteinizing hormone on their bioactivity in vitro, radioreceptor activity and immunoactivity.
To establish whether sialic acid content is responsible for an observed 7-8-fold variability in bioactivity in vitro of highly purified human pituitary luteinizing hormone (hLH) isoforms, the bioactivity in vitro, radioreceptor activity and immunoactivity of hLH isoforms were determined before and after enzymatic desialylation. Three immunofluorometric assays with different hLH specificities allowed characterization of 13-24 pituitary hLH isoform preparations of pI 7.03-8.98 in terms of sialic acid content (1-5 sialic acid residues per LH molecule), bioactivity in vitro (4030-30,000 I.U. mg[-1]), radioreceptor activity (6420-25,400 I.U. mg[-1]) and hLH immunoactivity (2900-4400 to 18,300-27,300 I.U. mg[-1]). Significant positive correlations between sialic acid content and either immunoactivity or in vitro bioactivity were observed, whereas radioreceptor activity showed a curvilinear response. Following more than 90% removal of sialic acid, both in vitro bioactivity and radioreceptor activity were increased, although specific activity still differed between isoforms; immunoactivities were unaffected. It is concluded that the presence of the sialic acid residue(s) on hLH isoforms does partially contribute to the in vitro bioactivity and radioreceptor activity of the isoforms, but that hLH immunoactivity is independent of sialic acid content.
Burgon
P G
PG
Prince Henry's Institute of Medical Research, Monash Medical Centre, Clayton, Vic., Australia.
Stanton
P G
PG
Pettersson
K
K
Robertson
D M
DM
D000818:Animals; D001681:Biological Assay; D004591:Electrophoresis, Polyacrylamide Gel; D015200:Fluoroimmunoassay; D006801:Humans; D007986:Luteinizing Hormone / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D019158:N-Acetylneuraminic Acid / Q000032:analysis*; D009439:Neuraminidase / Q000378:metabolism; D011869:Radioligand Assay; D012680:Sensitivity and Specificity
D007986:Luteinizing Hormone; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid