ABSTRACT
Human FSH consists of a mixture of isoforms that can be separated on the basis of differences in negative charge conferred by variations in the numbers of sialic acid residues that terminate oligosaccharide branches. Western analysis of human FSH isoforms separated by chromatofocusing revealed the presence of two human FSHbeta isoforms that differed in size. A low mol wt human FSHbeta isoform was associated with all FSH isoform fractions. A high mol wt human FSHbeta isoform was associated with the more acidic fractions and increased in relative abundance as the pI decreased. Characterization of representative human FSHbeta isoforms by mass spectrometry and automated Edman degradation revealed a low mol wt isoform that was not glycosylated. A high mol wt isoform was N-glycosylated at Asn residues 7 and 24. These results indicate that pituitary human FSH consists of two classes of molecules: those that possess a nonglycosylated beta-subunit and those that possess a glycosylated beta-subunit. Glycoprotein hormones are known to be elliptical molecules, and the beta-subunit oligosaccharides project outward from the short diameter, thereby increasing it. It is interesting to speculate that this change in shape might affect ultrafiltration rates, leading to differences in delivery rates to target tissues and elimination by filtration in the kidney.
Subject(s)
Follicle Stimulating Hormone/chemistry , Animals , Blotting, Western , Chorionic Gonadotropin/metabolism , Chromatography, Affinity , Diethylstilbestrol/pharmacology , Electrophoresis, Polyacrylamide Gel , Female , Follicle Stimulating Hormone/isolation & purification , Follicle Stimulating Hormone/pharmacology , Follicle Stimulating Hormone, beta Subunit , Glycosylation , Granulosa Cells/drug effects , Granulosa Cells/physiology , Humans , Immunoradiometric Assay , Male , Molecular Weight , Protein Isoforms/chemistry , Protein Isoforms/isolation & purification , Protein Isoforms/pharmacology , Radioligand Assay , Rats , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Testis/metabolismSubject(s)
Cholinesterase Inhibitors/toxicity , Cyprinidae/physiology , Cyprinodontiformes/physiology , Hexachlorocyclohexane/toxicity , Insecticides/toxicity , Parathion/toxicity , Animals , Biotransformation , Cholinesterase Inhibitors/metabolism , Circadian Rhythm , Female , Hexachlorocyclohexane/metabolism , Insecticides/metabolism , Lethal Dose 50 , Parathion/metabolism , Species SpecificityABSTRACT
Transforming growth factor-beta (TGF-beta), an ubiquitous regulatory peptide, has diverse effects on the differentiation and behavior of vascular smooth muscle cells (VSMC). However, the molecular mechanism through which TGF-alpha exerts its effects remains obscure. We investigated the phosphoinositide/protein kinase C [PKC] signaling pathway in the action of TGF-beta on cultured embryonic avian VSMC of differing lineage: a) thoracic aorta, derived from the neural crest; and b) abdominal aorta, derived from mesenchyme. The second messenger responsible for activation of PKC is sn-1,2-diacylglycerol [DAG]; TGF-beta increased the mass amounts of DAG in the membranes of neural crest-derived VSMC concurrent with translocation of PKC from the soluble to the membrane fraction, but TGF-beta had no effect on the DAG or PKC of mesenchyme-derived VSMC. TGF-beta potentiated the growth of platelet-derived growth factor (PDGF)-treated, neural crest-derived VSMC; but abolished PDGF-induced growth of mesenchymal cells. It is concluded that molecular and functional responses of VSMC to TGF-beta are heterogeneous and are functions of the embryonic lineage of the VSMC.
