Category Archives: Hormone - Part 3

Hormone-Specific Inhibitory Influence: RESULTS(6)

Despite the fact that it was possible to separate eFSHp hybrid dimers from unassociated subunits, the activities of the eFSHa:eFSHp hybrid preparation ranged from 43-58% that of the intact eFSH preparation in the CHO-hFSHR assay and chicken assay, respectively, to … Continue reading

Hormone-Specific Inhibitory Influence: RESULTS(5)

The magnitude of the difference in FSH receptor-binding activities due to the hormone-specific a-subunit influence was less for FSHp hybrids than for eLHp and eCGp hybrids. Selective removal of aAsn56 oligosaccharide increased the FSH receptor-binding activity of each hybrid and … Continue reading

Hormone-Specific Inhibitory Influence: RESULTS(4)

Comparison of the FSH receptor-binding activities of all intact and N56dg-a hybrid preparations was possible only on an analytical scale because of limited availability of hormone preparations from which subunit preparations were derived. However, it was an essential part of … Continue reading

Hormone-Specific Inhibitory Influence: RESULTS(3)

The increase in recombination efficiency of eCG(3 hybrid preparations following Asn56 deglycosylation appeared much lower than that obtained with eLH|3 hybrid preparations, and lower than that of eFSH|3 hybrid preparations, despite the fact that the molar ratio of a/p subunits … Continue reading

Hormone-Specific Inhibitory Influence: RESULTS(2)

The mass spectrometry data also indicated that eLHa possessed the smallest oligosaccharides attached to Asn56, 401 mass units smaller than those of eFSHa, and 845 mass units smaller than those of eCGa. Diagrammatic summaries of Asn56 oligosaccharide structural data including … Continue reading

Hormone-Specific Inhibitory Influence: RESULTS(1)

As indicated by the electrophoretic patterns for the eLHa, eFSHa, and eCGa preparations before and after PNGase digestion following SDS-PAGE under reducing conditions on a 15% polyacrylamide gel (Fig. 1), Asn56 carbohydrate was completely removed and inactivating proteolytic nicks were … Continue reading

Hormone-Specific Inhibitory Influence: MATERIALS AND METHODS(8)

Granulosa Cell Assay FSH bioassay using diethylstilbestrol (DES)-primed granulosa cells was carried out as previously described . Immature (20 to 21 days old) female Sprague-Dawley rats were implanted with 1-inch silicone elastomer capsules containing DES, and granulosa cells were harvested … Continue reading

Hormone-Specific Inhibitory Influence: MATERIALS AND METHODS(7)

The hFSHR was also included as it shows intermediate specificity and very high specific binding of 125I-eFSH (3040% of total cpm added). We observed the same phenomenon with all three receptors; this demonstrated that our results are not restricted to … Continue reading

Hormone-Specific Inhibitory Influence: MATERIALS AND METHODS(6)

This approach was taken as amino acid analysis has proven to be a highly unreliable method for quantification of these highly glycosylated glycoproteins, especially eCG . Highly purified eFSH was labeled with 125I using the chloramine T technique to specific … Continue reading

Hormone-Specific Inhibitory Influence: MATERIALS AND METHODS(5)

a-Subunit Asn56 Deglycosylation with Peptide-N-Glycosidase (PNGase) Selective Asn56 deglycosylation of 80-jxg samples of eLHa, eFSHa, or eCGa preparations was performed according the protocol described recently . Preparative-scale Asn56 deglycosylation was carried out on 1-mg samples of eLHa, eFSHa, or eCGa … Continue reading

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