Naturally Occurring Follicle-Stimulating Hormone Glycosylation Variants

Davis, John S.; Kumar, T. Rajendra; May, Jeffrey V.; Bousfield, George R.

doi:10.4172/2153-0637.1000e117

Cited by 35 publications

(25 citation statements)

References 69 publications

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“…The existence of macro-heterogeneity in FSH preparations contributed by FSHβ subunits differing in N-glycosylation was recently confirmed [4-6, 14, 17-19]. Four FSH glycoforms were identified by direct purification from pituitary extracts [4-7, 14, 17-19] and following recombinant expression of genes encoding specific N-glycosylation mutant FSHβ subunits [4-7, 14, 17-19].…”

Section: Discussionmentioning

confidence: 99%

“…It was predicted that such age-related FSH glycoforms act differently in different target cells [4, 14, 17]. Direct purification of FSH glycoforms from human pituitaries and/or urine collected at different ages and their characterization indeed identified hypo-glycosylated FSH is a mixture of FSH 18 and FSH 21 glycoforms present as the dominant form in young age and fully-glycosylated FSH, designated as FSH 24 is the dominant form in old age [4, 6, 14, 17-20]. Recombinant expression in a heterologous GH 3 cell culture system followed by purification and biochemical characterization revealed that these FSH glycoforms differ in the number or location of N-linked glycan chains on FSHβ subunit [4-7, 14].…”

Section: Introductionmentioning

confidence: 99%

“…Direct purification of FSH glycoforms from human pituitaries and/or urine collected at different ages and their characterization indeed identified hypo-glycosylated FSH is a mixture of FSH 18 and FSH 21 glycoforms present as the dominant form in young age and fully-glycosylated FSH, designated as FSH 24 is the dominant form in old age [4, 6, 14, 17-20]. Recombinant expression in a heterologous GH 3 cell culture system followed by purification and biochemical characterization revealed that these FSH glycoforms differ in the number or location of N-linked glycan chains on FSHβ subunit [4-7, 14]. While both N-linked glycans are present at Asn 7 and Asn 24 in the FSHβ subunit of fully-glycosylated FSH 24 , a single N-linked glycan is present only at Asn 7 in the FSHβ subunit of FSH 21 or at Asn 24 in the FSHβ subunit of FSH 18 .…”

Section: Introductionmentioning

confidence: 99%

“…While both N-linked glycans are present at Asn 7 and Asn 24 in the FSHβ subunit of fully-glycosylated FSH 24 , a single N-linked glycan is present only at Asn 7 in the FSHβ subunit of FSH 21 or at Asn 24 in the FSHβ subunit of FSH 18 . The latter two glycoforms are collectively known as hypo-glycosylated FSH and referred to as FSH 21/18 [4-7, 14, 19]. …”

Section: Introductionmentioning

confidence: 99%

“…Both the pituitary-derived preparations and GH 3 cell-derived recombinant FSH glycoforms have been biochemically characterized in vitro and compared to fully-glycosylated FSH 24 with regard to receptor binding and dissociation kinetics, glycan composition by mass spectrometry and in vitro bioactivity using radio-receptor assays [4-7, 14, 18, 19]. More recently, these FSH glycoforms have also been tested in an FSH-responsive KGN granulosa cell line [17].…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of in vivo bioactivities of recombinant hypo- (FSH 21/18 ) and fully- (FSH 24 ) glycosylated human FSH glycoforms in Fshb null mice

Wang

May

Butnev

et al. 2016

Molecular and Cellular Endocrinology

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The hormone - specific FSHβ subunit of the human FSH heterodimer consists of N-linked glycans at Asn7 and Asn24 residues that are co-translationally attached early during subunit biosynthesis. Differences in the number of N-glycans (none, one or two) on the human FSHβ subunit contribute to macroheterogeneity in the FSH heterodimer. The resulting FSH glycoforms are termed hypo-glycosylated (FSH21/18, missing either an Asn24 or Asn7 N-glycan chain on the β - subunit, respectively) or fully glycosylated (FSH24, possessing of both Asn7 and Asn24 N-linked glycans on the β - subunit) FSH. The recombinant versions of human FSH glycoforms (FSH21/18 and FSH24) have been purified and biochemically characterized. In vitro functional studies have indicated that FSH21/18 exhibits faster FSH- receptor binding kinetics and is much more active than FSH24 in every assay tested to date. However, the in vivo bioactivity of the hypoglycosylated FSH glycoform has never been tested. Here, we evaluated the in vivo bioactivities of FSH glycoforms in Fshb null mice using a pharmacological rescue approach. In Fshb null female mice, both hypo- and fully-glycosylated FSH elicited an ovarian weight gain response by 48h and induced ovarian genes in a dose- and time-dependent manner. Quantification by real time qPCR assays indicated that hypo-glycosylated FSH21/18 was bioactive in vivo and induced FSH-responsive ovarian genes similar to fully-glycosylated FSH24. Western blot analyses followed by densitometry of key signaling components downstream of the FSH-receptor confirmed that the hypo-glycosylated FSH21/18 elicited a response similar to that by fully-glycosylated FSH24 in ovaries of Fshb null mice. When injected into Fshb null males, hypo-glycosylated FSH21/18 was more active than the fully-glycosylated FSH24 in inducing FSH-responsive genes and Sertoli cell proliferation. Thus, our data establish that recombinant hypo-glycosylated human FSH21/18 glycoform elicits bioactivity in vivo similar to the fully-glycosylated FSH. Our studies may have clinical implications particularly in formulating FSH-based ovarian follicle induction protocols using a combination of different human FSH glycoforms.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Evaluation of in vivo bioactivities of recombinant hypo- (FSH 21/18 ) and fully- (FSH 24 ) glycosylated human FSH glycoforms in Fshb null mice

Wang

May

Butnev

et al. 2016

Molecular and Cellular Endocrinology

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FGR (Gene Name)

Berton¹,

Baruzzi²

2018

Encyclopedia of Signaling Molecules

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(Duh et al. 1994; Holthuis et al. 1994). The sequence analyses revealed that fascin proteins form a unique family of actin-bundling proteins, sharing no apparent homology with nonfascin actin-bundling proteins including alpha-actinin, villin, and fimbrin.Vertebrates have three fascin genes (fascin-1 through À3): fascin-1 shows widespread expression in a variety of tissues, whereas expression of fascin-2 is restricted to retina and hair cell stereocilia, whereas fascin-3 is restricted to testis. Fascin is absent in Dictyostelium discoideum, C. elegans, or yeasts. As expected from the actin-bundling activity, fascin is mainly localized in filopodia of mammalian cultured cells, microvilli of sea urchin eggs, and drosophila bristles. Consistent with highly motile structure of filopodia, fascin is shown to promote cell motility and metastasis of tumor cells. Intriguingly, a fascin homologue is found in microvilli and filopodia of choanoflagellates. Because this unicellular organism is considered as the last ancestor of multicellular animals, fascin1 is suggested to be an ancestral component for filopodial assembly machinery (Sebe-Pedros et al. 2013). Structure and Function of FascinX-ray structural analyses revealed that fascin has a b-trefoil structure (Sedeh et al. 2010), showing Fascin1 plays an important role in the formation of filopodia in mammalian cells. In vitro, fascin1 makes parallel actin bundles with uniform polarity, which is consistent with its localization in filopodia. Upregulation of fascin1 induces membrane protrusions and increases cell motility of epithelial cells, as well as colonic epithelial and carcinoma cells. Conversely, Fascin1 knock down has been reported to block filopodia assembly of B16F1 mouse melanoma cells, as well as colon carcinoma cells and mature antigen-presenting dendritic cells (DCs) (Ross et al. 1998).In addition to binding to actin filaments, fascin1 has been recently reported to bind directly to microtubules (Villari et al. 2015). Fascin1-microtubule binding occurred independently of fascin1-actin binding. The association was shown to increase the dynamics of focal adhesions, as well as that of microtubules, thereby controlling cell motility. This regulation of focal adhesion dynamics may be controlled via FAK because fascin1 was found to bind to FAK. Genetic AnalysesAnalyses of Drosophila singed mutations indicate that fascin1 is involved in female sterility, in addition to the gnarled bristle phenotype. In Drosophila oogenesis, each developing oocyte is surrounded by and connected to 15 nurse cells via intercellular bridges. Nurse cell cytoplasmic contents flow into the oocyte along actin filaments traversing these cytoplasmic bridges. A singed allele affects the microfilament structure required for this nurse cell cytoplasmic flow, resulting in female sterility (Cant et al. 1994). Interestingly, fascin1 is involved in prostaglandin-mediated actin remodeling (Groen et al. 2012). Prostaglandin is synthesized in Drosophila by peroxidase (Pxt), a cyclooxygenase (COX)-like...

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FGF (Fibroblast Growth Factor)

Katoh¹

2018

Encyclopedia of Signaling Molecules

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Naturally Occurring Follicle-Stimulating Hormone Glycosylation Variants

Cited by 35 publications

References 69 publications

Evaluation of in vivo bioactivities of recombinant hypo- (FSH 21/18 ) and fully- (FSH 24 ) glycosylated human FSH glycoforms in Fshb null mice

Evaluation of in vivo bioactivities of recombinant hypo- (FSH 21/18 ) and fully- (FSH 24 ) glycosylated human FSH glycoforms in Fshb null mice

FGR (Gene Name)

FGF (Fibroblast Growth Factor)

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