Biological and immunological activity in bovine luteinizing hormone charge isoforms

Abstract

Luteinizing hormone (LH) undergoes posttranslational modifications that originate different charge isoforms. The study evaluated the differences in biological (B) and immunological (I) activity between isoforms of bovine LH. Isoforms were isolated by chromatofocusing from anterior pituitary glycoprotein extract. The biological activity was evaluated in an in vitro bioassay. Immunological activity was measured with a radioimmunoassay (RIA) specific for LH. The USDA-bLH-B5 standard was utilized as the reference. LH isoforms were grouped by their pH range of elution, in basic (A, pH, 10.75-9.75; B, pH 9.58-8.41), neutral (C pH, 7.98-6.89) and acidic (D, pH, 6.88-5.41; E, pH 5.36-3.46).

The molecular weight of the heterodimer of each isoform was similar to the LH standard, estimated to be 36.5 kDa. Immunological and biological activity behaved in a dose-dependent manner. With respect to the LH standard, all isoforms required higher protein concentration to reach the IC50 of the inhibition curve. In the bioassay EC50 value for cAMP production was significantly different among isoforms; the neutral isoform showed a lower EC50 which was interpreted as more bioactive, the acidic isoform E showed an EC50 being the least bioactive and the basic was intermediate (P<0.05). In conclusion, the results suggest a quantitative effect of LH charge isoforms on the cAMP production, per unit of immunoreactive LH in the bioassay.