Hypo-glycosylated human follicle-stimulating hormone (hFSH21/18) is much more active in vitro than fully-glycosylated hFSH (hFSH24)

George R. Bousfield, Vladimir Y. Butnev, Viktor Y. Butnev, Yasuaki Hiromasa, David J. Harvey, Jeffrey V. May

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Hypo-glycosylated hFSH21/18 (possesses FSHβ21 and FSHβ18bands) was isolated from hLH preparations by immunoaffinity chromatography followed by gel filtration. Fully-glycosylated hFSH24 was prepared by combining the fully-glycosylated FSHβ24 variant with hCGα and isolating the heterodimer. The hFSH21/18 glycoform preparation was significantly smaller than the hFSH24 preparation and possessed 60% oligomannose glycans, which is unusual for hFSH. Hypo-glycosylated hFSH21/18 was 9- to 26-fold more active than fully-glycosylated hFSH24 in FSH radioligand assays. Significantly greater binding of 125I-hFSH21/18 tracer than hFSH24 tracer was observed in all competitive binding assays. In addition, higher binding of hFSH21/18 was noted in association and saturation binding assays, in which twice as much hFSH21/18 was bound as hFSH24. This suggests that more ligand binding sites are available to hFSH21/18 in FSHR than to hFSH24. Hypo-glycosylated hFSH21/18 also bound rat FSHRs more rapidly, exhibiting almost no lag in binding, whereas hFSH24 specific binding proceeded very slowly for almost the first hour of incubation.

Original languageEnglish
Pages (from-to)989-997
Number of pages9
JournalMolecular and Cellular Endocrinology
Volume382
Issue number2
DOIs
Publication statusPublished - Feb 15 2014

Fingerprint

Human Follicle Stimulating Hormone
Radioligand Assay
Competitive Binding
Gel Chromatography
Polysaccharides
Assays
Binding Sites
Ligands
Chromatography
Rats
Gels
Association reactions
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Endocrinology

Cite this

Hypo-glycosylated human follicle-stimulating hormone (hFSH21/18) is much more active in vitro than fully-glycosylated hFSH (hFSH24). / Bousfield, George R.; Butnev, Vladimir Y.; Butnev, Viktor Y.; Hiromasa, Yasuaki; Harvey, David J.; May, Jeffrey V.

In: Molecular and Cellular Endocrinology, Vol. 382, No. 2, 15.02.2014, p. 989-997.

Research output: Contribution to journalArticle

Bousfield, George R. ; Butnev, Vladimir Y. ; Butnev, Viktor Y. ; Hiromasa, Yasuaki ; Harvey, David J. ; May, Jeffrey V. / Hypo-glycosylated human follicle-stimulating hormone (hFSH21/18) is much more active in vitro than fully-glycosylated hFSH (hFSH24). In: Molecular and Cellular Endocrinology. 2014 ; Vol. 382, No. 2. pp. 989-997.
@article{84d1021215c143eea9940167a980cb00,
title = "Hypo-glycosylated human follicle-stimulating hormone (hFSH21/18) is much more active in vitro than fully-glycosylated hFSH (hFSH24)",
abstract = "Hypo-glycosylated hFSH21/18 (possesses FSHβ21 and FSHβ18bands) was isolated from hLH preparations by immunoaffinity chromatography followed by gel filtration. Fully-glycosylated hFSH24 was prepared by combining the fully-glycosylated FSHβ24 variant with hCGα and isolating the heterodimer. The hFSH21/18 glycoform preparation was significantly smaller than the hFSH24 preparation and possessed 60{\%} oligomannose glycans, which is unusual for hFSH. Hypo-glycosylated hFSH21/18 was 9- to 26-fold more active than fully-glycosylated hFSH24 in FSH radioligand assays. Significantly greater binding of 125I-hFSH21/18 tracer than hFSH24 tracer was observed in all competitive binding assays. In addition, higher binding of hFSH21/18 was noted in association and saturation binding assays, in which twice as much hFSH21/18 was bound as hFSH24. This suggests that more ligand binding sites are available to hFSH21/18 in FSHR than to hFSH24. Hypo-glycosylated hFSH21/18 also bound rat FSHRs more rapidly, exhibiting almost no lag in binding, whereas hFSH24 specific binding proceeded very slowly for almost the first hour of incubation.",
author = "Bousfield, {George R.} and Butnev, {Vladimir Y.} and Butnev, {Viktor Y.} and Yasuaki Hiromasa and Harvey, {David J.} and May, {Jeffrey V.}",
year = "2014",
month = "2",
day = "15",
doi = "10.1016/j.mce.2013.11.008",
language = "English",
volume = "382",
pages = "989--997",
journal = "Molecular and Cellular Endocrinology",
issn = "0303-7207",
publisher = "Elsevier Ireland Ltd",
number = "2",

}

TY - JOUR

T1 - Hypo-glycosylated human follicle-stimulating hormone (hFSH21/18) is much more active in vitro than fully-glycosylated hFSH (hFSH24)

AU - Bousfield, George R.

AU - Butnev, Vladimir Y.

AU - Butnev, Viktor Y.

AU - Hiromasa, Yasuaki

AU - Harvey, David J.

AU - May, Jeffrey V.

PY - 2014/2/15

Y1 - 2014/2/15

N2 - Hypo-glycosylated hFSH21/18 (possesses FSHβ21 and FSHβ18bands) was isolated from hLH preparations by immunoaffinity chromatography followed by gel filtration. Fully-glycosylated hFSH24 was prepared by combining the fully-glycosylated FSHβ24 variant with hCGα and isolating the heterodimer. The hFSH21/18 glycoform preparation was significantly smaller than the hFSH24 preparation and possessed 60% oligomannose glycans, which is unusual for hFSH. Hypo-glycosylated hFSH21/18 was 9- to 26-fold more active than fully-glycosylated hFSH24 in FSH radioligand assays. Significantly greater binding of 125I-hFSH21/18 tracer than hFSH24 tracer was observed in all competitive binding assays. In addition, higher binding of hFSH21/18 was noted in association and saturation binding assays, in which twice as much hFSH21/18 was bound as hFSH24. This suggests that more ligand binding sites are available to hFSH21/18 in FSHR than to hFSH24. Hypo-glycosylated hFSH21/18 also bound rat FSHRs more rapidly, exhibiting almost no lag in binding, whereas hFSH24 specific binding proceeded very slowly for almost the first hour of incubation.

AB - Hypo-glycosylated hFSH21/18 (possesses FSHβ21 and FSHβ18bands) was isolated from hLH preparations by immunoaffinity chromatography followed by gel filtration. Fully-glycosylated hFSH24 was prepared by combining the fully-glycosylated FSHβ24 variant with hCGα and isolating the heterodimer. The hFSH21/18 glycoform preparation was significantly smaller than the hFSH24 preparation and possessed 60% oligomannose glycans, which is unusual for hFSH. Hypo-glycosylated hFSH21/18 was 9- to 26-fold more active than fully-glycosylated hFSH24 in FSH radioligand assays. Significantly greater binding of 125I-hFSH21/18 tracer than hFSH24 tracer was observed in all competitive binding assays. In addition, higher binding of hFSH21/18 was noted in association and saturation binding assays, in which twice as much hFSH21/18 was bound as hFSH24. This suggests that more ligand binding sites are available to hFSH21/18 in FSHR than to hFSH24. Hypo-glycosylated hFSH21/18 also bound rat FSHRs more rapidly, exhibiting almost no lag in binding, whereas hFSH24 specific binding proceeded very slowly for almost the first hour of incubation.

UR - http://www.scopus.com/inward/record.url?scp=84890612296&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84890612296&partnerID=8YFLogxK

U2 - 10.1016/j.mce.2013.11.008

DO - 10.1016/j.mce.2013.11.008

M3 - Article

VL - 382

SP - 989

EP - 997

JO - Molecular and Cellular Endocrinology

JF - Molecular and Cellular Endocrinology

SN - 0303-7207

IS - 2

ER -