Determination of the Disulfide Structure andN-Glycosylation Sites of the Extracellular Domain of the Human Signal Transducer gp130

Moritz, Robert L.; Hall, Nathan E.; Connolly, Lisa; Simpson, Richard J.

doi:10.1074/jbc.m009979200

Cited by 28 publications

(30 citation statements)

References 78 publications

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“…4C). These data confirmed the findings of Moritz et al (21) and indicated that the gp130 part of sgp130Fc-dNG was indeed free of N-glycans.…”

Section: Enzymatic Analysis Of N-and O-glycosylation Of Sgp130fc-dng-supporting

confidence: 82%

“…In 2001, Moritz et al (21) 224 and Asn 368 were not. Of these two sites, N-glycosylation on Asn 224 is practically impossible, as Asn 224 is followed by a proline and is also partially buried in the gp130 structure (21,22). The crystal structure of the complete extracellular part of gp130 is still unknown, but the structure of the three ligandbinding domains D1-D3 or D2 ϩ D3 has been solved in complex with viral IL-6 (23), LIF (24), or IL-6⅐sIL-6R (25).…”

mentioning

confidence: 99%

“…gp130 belongs to the class of "tall cytokine receptors" that feature three fibronectin-type III-like domains between their ligand-binding domains and their transmembrane domain (20). In 2001, Moritz et al (21) 224 and Asn 368 were not. Of these two sites, N-glycosylation on Asn 224 is practically impossible, as Asn 224 is followed by a proline and is also partially buried in the gp130 structure (21,22).…”

mentioning

confidence: 99%

“…The crystal structure of the complete extracellular part of gp130 is still unknown, but the structure of the three ligandbinding domains D1-D3 or D2 ϩ D3 has been solved in complex with viral IL-6 (23), LIF (24), or IL-6⅐sIL-6R (25). Two of these studies used gp130 fragments produced in insect cells in the presence of tunicamycin as a global N-glycosylation inhibitor (23,24), whereas the third study employed a gp130 expression construct in which Asn 21 , Asn 109 , Asn 135 , Asn 205 , and Asn 224 (but not Asn 61 ) of gp130 were mutated to glutamine (25). Both tunicamycin and the Asn/Gln mutations resulted in reduced yield of the protein in insect cells but did not change its biochemical behavior or binding properties (23)(24)(25).…”

mentioning

confidence: 99%

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N-Linked Glycosylation Is Essential for the Stability but Not the Signaling Function of the Interleukin-6 Signal Transducer Glycoprotein 130

Waetzig

Chalaris

Rosenstiel³

et al. 2010

Journal of Biological Chemistry

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N-Linked glycosylation is an important determinant of protein structure and function. The interleukin-6 signal transducer glycoprotein 130 (gp130) is a common co-receptor for cytokines of the interleukin (IL)-6 family and is N-glycosylated at 9 of 11 potential sites. Whereas N-glycosylation of the extracellular domains D1-D3 of gp130 has been shown to be dispensable for binding of the gp130 ligand IL-6 and its cognate receptor in vitro, the role of the N-linked glycans on domains D4 and D6 is still unclear. We have mutated the asparagines of all nine functional N-glycosylation sites of gp130 to glutamine and systematically analyzed the consequences of deleted N-glycosylation (dNG) in both cellular gp130 and in a soluble gp130-IgG1-Fc fusion protein (sgp130Fc). Our results show that sgp130Fc-dNG is inherently unstable and degrades rapidly under conditions that do not harm wild-type sgp130Fc. Consistently, the bulk of cellular gp130-dNG is not transported to the plasma membrane but is degraded in the proteasome. However, the small quantities of gp130-dNG, which do reach the cell surface, are still able to activate the key gp130 signaling target signal transducer and activator of transcription-3 (STAT3) upon binding of the agonistic complex of IL-6 and soluble IL-6 receptor. In conclusion, N-linked glycosylation is required for the stability but not the signal-transducing function of gp130.

show abstract

“…4C). These data confirmed the findings of Moritz et al (21) and indicated that the gp130 part of sgp130Fc-dNG was indeed free of N-glycans.…”

Section: Enzymatic Analysis Of N-and O-glycosylation Of Sgp130fc-dng-supporting

confidence: 82%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

N-Linked Glycosylation Is Essential for the Stability but Not the Signaling Function of the Interleukin-6 Signal Transducer Glycoprotein 130

Waetzig

Chalaris

Rosenstiel³

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Several orientations of these domains are possible, but in the absence of any structural information, they are difficult to dock with sufficient surety. By using homology models of these three domains, one can orient in such a way so they contact Cluster 3 on D 3 of IL-6R and form a disulfide link proposed (52) between D 5 of gp130 underneath the complex, as shown schematically in Fig. 6b.…”

Section: Resultsmentioning

confidence: 99%

Structure of the extracellular domains of the human interleukin-6 receptor α-chain

Varghese

Moritz

Lou

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

114

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Thiol‐Disulfide Oxidoreduction of Protein Cysteines: Old Methods Revisited for Proteomics

Bonetto

Ghezzi

2006

Redox Proteomics

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Determination of the Disulfide Structure andN-Glycosylation Sites of the Extracellular Domain of the Human Signal Transducer gp130

Cited by 28 publications

References 78 publications

N-Linked Glycosylation Is Essential for the Stability but Not the Signaling Function of the Interleukin-6 Signal Transducer Glycoprotein 130

N-Linked Glycosylation Is Essential for the Stability but Not the Signaling Function of the Interleukin-6 Signal Transducer Glycoprotein 130

Structure of the extracellular domains of the human interleukin-6 receptor α-chain

Thiol‐Disulfide Oxidoreduction of Protein Cysteines: Old Methods Revisited for Proteomics

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