Characterization of human FSH isoforms reveals a nonglycosylated beta-subunit in addition to the conventional glycosylated beta-subunit.
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.
Walton
W J
WJ
Department of Biological Sciences, Wichita State University, Wichita, Kansas 67260-0026, USA.
Nguyen
V T
VT
Butnev
V Y
VY
Singh
V
V
Moore
W T
WT
Bousfield
G R
GR
D000818:Animals; D015153:Blotting, Western; D006063:Chorionic Gonadotropin / Q000378:metabolism; D002846:Chromatography, Affinity; D004054:Diethylstilbestrol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005640:Follicle Stimulating Hormone / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D037201:Follicle Stimulating Hormone, beta Subunit; D006031:Glycosylation; D006107:Granulosa Cells / Q000187:drug effects / Q000502:physiology; D006801:Humans; D015592:Immunoradiometric Assay; D008297:Male; D008970:Molecular Weight; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D011869:Radioligand Assay; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013737:Testis / Q000378:metabolism
D006063:Chorionic Gonadotropin; D037201:Follicle Stimulating Hormone, beta Subunit; D020033:Protein Isoforms; D004054:Diethylstilbestrol; D005640:Follicle Stimulating Hormone