Deglycosylated human follitropin: characterization and effects on adenosine cyclic 3',5'-phosphate production in porcine granulosa cells.
Chemical deglycosylation of gonadotropins with hydrogen fluoride (HF) has facilitated the investigation of the structure-function relationship of the individual peptide and oligosaccharide moieties in the mechanism of hormone action. These studies have dealt almost exclusively with lutropin or human choriogonadotropin. We report here the chemical characterization and biological properties of deglycosylated human follitropin (degly hFSH). Results indicate that deglycosylation of hFSH by HF removes 89% of the total carbohydrate without disruption of the peptide chain or significant loss of amino acid residues. However, a change in the conformation of the molecule was observed by measurement of the far-ultraviolet circular dichroic spectrum. The degly hFSH showed a 44% reduction in binding when tested in a FSH radioimmunoassay utilizing a polyclonal antibody. Binding of the degly hFSH to FSH-responsive tissues showed that the altered hormone bound with equal or better avidity than the intact hormone while the association constants were approximately the same for both preparations. The degly hFSH alone did not stimulate the FSH-stimulatable adenylyl cyclase (AC) activity of cellular homogenates of small follicle porcine granulosa cells. Furthermore, degly hFSH was a potent antagonist of hFSH-stimulatable AC activity when coincubated with intact hFSH. In intact granulosa cells, both the hFSH and the degly hFSH stimulated cAMP production and release by these cells. However, the degly hFSH was one-tenth as effective as the intact hormone.(ABSTRACT TRUNCATED AT 250 WORDS)
D000262:Adenylyl Cyclases / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D002942:Circular Dichroism; D003429:Cross Reactions; D000242:Cyclic AMP / Q000378:metabolism*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006858:Hydrofluoric Acid; D007700:Kinetics; D008297:Male; D011863:Radioimmunoassay; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism; D011962:Receptors, FSH; D013552:Swine; D013737:Testis / Q000378:metabolism
D000596:Amino Acids; D002241:Carbohydrates; D011956:Receptors, Cell Surface; D011962:Receptors, FSH; C049669:follicle stimulating hormone, deglycosylated; D005640:Follicle Stimulating Hormone; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D006858:Hydrofluoric Acid