Figure 2.

A negative effect of the prodomain on AMH complex binding to AMHR2 is observed when AMHR2 is on a surface but not in solution.A and B, ELISA format and results for comparing binding of c-AMH or GF to AMHR2 when the receptor is on a surface. The AMHR2-Fc/Fc was captured on a goat antihuman Fc Ab and incubated with various concentrations of AMH; bound AMH was detected with biotinylated anti-C-terminal AMH mAb-C₁ and streptavidin conjugated to horse radish peroxidase (HRP). C and D, ELISA format and results for comparing binding of c-AMH or GF to AMHR2 when the receptor is presented in solution. AMH was captured on mAb-C₁ and incubated with various concentrations of AMHR2-Fc/Fc; bound AMHR2-Fc/Fc was detected with an antihuman-Fc Ab conjugated to HRP. E and F, ELISA format and results for assessing whether AMHR2-Fc/Fc in solution can induce displacement of the prodomain. c-AMH, after capture on mAb-C₁, was incubated with AMHR2-Fc/Fc (33 nM) or buffer for 2 h, prior to detection with mAb-N₁. Data in B and D were fit to the quadratic equation for a reversible association reaction. Error bars show standard deviations. Multivalent interactions are not shown in the diagrams depicting ELISA formats. Ab, antibody; AMH, anti-Müllerian hormone; AMHR2, anti-Müllerian hormone receptor type-2; c-AMH, cleaved AMH; GF, growth factor.