Glycosylation Tunes Neuroserpin Physiological and Pathological Properties

Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type...
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Autor*in:	Cristina Visentin [verfasserIn] Luca Broggini [verfasserIn] Benedetta Maria Sala [verfasserIn] Rosaria Russo [verfasserIn] Alberto Barbiroli [verfasserIn] Carlo Santambrogio [verfasserIn] Simona Nonnis [verfasserIn] Anatoly Dubnovitsky [verfasserIn] Martino Bolognesi [verfasserIn] Elena Miranda [verfasserIn] Adnane Achour [verfasserIn] Stefano Ricagno [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2020

Schlagwörter:	neuroserpin protein polymerisation glycosylation

Übergeordnetes Werk:	In: International Journal of Molecular Sciences - MDPI AG, 2003, 21(2020), 9, p 3235
Übergeordnetes Werk:	volume:21 ; year:2020 ; number:9, p 3235

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc Journal toc

DOI / URN:	10.3390/ijms21093235

Katalog-ID:	DOAJ003398080

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520			\|a Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB.
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allfields	10.3390/ijms21093235 doi (DE-627)DOAJ003398080 (DE-599)DOAJb4f1f4887daf4073bafdfe98de5943d7 DE-627 ger DE-627 rakwb eng QH301-705.5 QD1-999 Cristina Visentin verfasserin aut Glycosylation Tunes Neuroserpin Physiological and Pathological Properties 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB. neuroserpin protein polymerisation glycosylation Biology (General) Chemistry Luca Broggini verfasserin aut Benedetta Maria Sala verfasserin aut Rosaria Russo verfasserin aut Alberto Barbiroli verfasserin aut Carlo Santambrogio verfasserin aut Simona Nonnis verfasserin aut Anatoly Dubnovitsky verfasserin aut Martino Bolognesi verfasserin aut Elena Miranda verfasserin aut Adnane Achour verfasserin aut Stefano Ricagno verfasserin aut In International Journal of Molecular Sciences MDPI AG, 2003 21(2020), 9, p 3235 (DE-627)316340715 (DE-600)2019364-6 14220067 nnns volume:21 year:2020 number:9, p 3235 https://doi.org/10.3390/ijms21093235 kostenfrei https://doaj.org/article/b4f1f4887daf4073bafdfe98de5943d7 kostenfrei https://www.mdpi.com/1422-0067/21/9/3235 kostenfrei https://doaj.org/toc/1661-6596 Journal toc kostenfrei https://doaj.org/toc/1422-0067 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 21 2020 9, p 3235
spelling	10.3390/ijms21093235 doi (DE-627)DOAJ003398080 (DE-599)DOAJb4f1f4887daf4073bafdfe98de5943d7 DE-627 ger DE-627 rakwb eng QH301-705.5 QD1-999 Cristina Visentin verfasserin aut Glycosylation Tunes Neuroserpin Physiological and Pathological Properties 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB. neuroserpin protein polymerisation glycosylation Biology (General) Chemistry Luca Broggini verfasserin aut Benedetta Maria Sala verfasserin aut Rosaria Russo verfasserin aut Alberto Barbiroli verfasserin aut Carlo Santambrogio verfasserin aut Simona Nonnis verfasserin aut Anatoly Dubnovitsky verfasserin aut Martino Bolognesi verfasserin aut Elena Miranda verfasserin aut Adnane Achour verfasserin aut Stefano Ricagno verfasserin aut In International Journal of Molecular Sciences MDPI AG, 2003 21(2020), 9, p 3235 (DE-627)316340715 (DE-600)2019364-6 14220067 nnns volume:21 year:2020 number:9, p 3235 https://doi.org/10.3390/ijms21093235 kostenfrei https://doaj.org/article/b4f1f4887daf4073bafdfe98de5943d7 kostenfrei https://www.mdpi.com/1422-0067/21/9/3235 kostenfrei https://doaj.org/toc/1661-6596 Journal toc kostenfrei https://doaj.org/toc/1422-0067 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 21 2020 9, p 3235
allfields_unstemmed	10.3390/ijms21093235 doi (DE-627)DOAJ003398080 (DE-599)DOAJb4f1f4887daf4073bafdfe98de5943d7 DE-627 ger DE-627 rakwb eng QH301-705.5 QD1-999 Cristina Visentin verfasserin aut Glycosylation Tunes Neuroserpin Physiological and Pathological Properties 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB. neuroserpin protein polymerisation glycosylation Biology (General) Chemistry Luca Broggini verfasserin aut Benedetta Maria Sala verfasserin aut Rosaria Russo verfasserin aut Alberto Barbiroli verfasserin aut Carlo Santambrogio verfasserin aut Simona Nonnis verfasserin aut Anatoly Dubnovitsky verfasserin aut Martino Bolognesi verfasserin aut Elena Miranda verfasserin aut Adnane Achour verfasserin aut Stefano Ricagno verfasserin aut In International Journal of Molecular Sciences MDPI AG, 2003 21(2020), 9, p 3235 (DE-627)316340715 (DE-600)2019364-6 14220067 nnns volume:21 year:2020 number:9, p 3235 https://doi.org/10.3390/ijms21093235 kostenfrei https://doaj.org/article/b4f1f4887daf4073bafdfe98de5943d7 kostenfrei https://www.mdpi.com/1422-0067/21/9/3235 kostenfrei https://doaj.org/toc/1661-6596 Journal toc kostenfrei https://doaj.org/toc/1422-0067 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 21 2020 9, p 3235
allfieldsGer	10.3390/ijms21093235 doi (DE-627)DOAJ003398080 (DE-599)DOAJb4f1f4887daf4073bafdfe98de5943d7 DE-627 ger DE-627 rakwb eng QH301-705.5 QD1-999 Cristina Visentin verfasserin aut Glycosylation Tunes Neuroserpin Physiological and Pathological Properties 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB. neuroserpin protein polymerisation glycosylation Biology (General) Chemistry Luca Broggini verfasserin aut Benedetta Maria Sala verfasserin aut Rosaria Russo verfasserin aut Alberto Barbiroli verfasserin aut Carlo Santambrogio verfasserin aut Simona Nonnis verfasserin aut Anatoly Dubnovitsky verfasserin aut Martino Bolognesi verfasserin aut Elena Miranda verfasserin aut Adnane Achour verfasserin aut Stefano Ricagno verfasserin aut In International Journal of Molecular Sciences MDPI AG, 2003 21(2020), 9, p 3235 (DE-627)316340715 (DE-600)2019364-6 14220067 nnns volume:21 year:2020 number:9, p 3235 https://doi.org/10.3390/ijms21093235 kostenfrei https://doaj.org/article/b4f1f4887daf4073bafdfe98de5943d7 kostenfrei https://www.mdpi.com/1422-0067/21/9/3235 kostenfrei https://doaj.org/toc/1661-6596 Journal toc kostenfrei https://doaj.org/toc/1422-0067 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 21 2020 9, p 3235
allfieldsSound	10.3390/ijms21093235 doi (DE-627)DOAJ003398080 (DE-599)DOAJb4f1f4887daf4073bafdfe98de5943d7 DE-627 ger DE-627 rakwb eng QH301-705.5 QD1-999 Cristina Visentin verfasserin aut Glycosylation Tunes Neuroserpin Physiological and Pathological Properties 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB. neuroserpin protein polymerisation glycosylation Biology (General) Chemistry Luca Broggini verfasserin aut Benedetta Maria Sala verfasserin aut Rosaria Russo verfasserin aut Alberto Barbiroli verfasserin aut Carlo Santambrogio verfasserin aut Simona Nonnis verfasserin aut Anatoly Dubnovitsky verfasserin aut Martino Bolognesi verfasserin aut Elena Miranda verfasserin aut Adnane Achour verfasserin aut Stefano Ricagno verfasserin aut In International Journal of Molecular Sciences MDPI AG, 2003 21(2020), 9, p 3235 (DE-627)316340715 (DE-600)2019364-6 14220067 nnns volume:21 year:2020 number:9, p 3235 https://doi.org/10.3390/ijms21093235 kostenfrei https://doaj.org/article/b4f1f4887daf4073bafdfe98de5943d7 kostenfrei https://www.mdpi.com/1422-0067/21/9/3235 kostenfrei https://doaj.org/toc/1661-6596 Journal toc kostenfrei https://doaj.org/toc/1422-0067 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 21 2020 9, p 3235
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callnumber-first	Q - Science
author	Cristina Visentin
spellingShingle	Cristina Visentin misc QH301-705.5 misc QD1-999 misc neuroserpin misc protein polymerisation misc glycosylation misc Biology (General) misc Chemistry Glycosylation Tunes Neuroserpin Physiological and Pathological Properties
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title_sort	glycosylation tunes neuroserpin physiological and pathological properties
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title_auth	Glycosylation Tunes Neuroserpin Physiological and Pathological Properties
abstract	Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB.
abstractGer	Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB.
abstract_unstemmed	Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. Thus, our results support the relevance of gNS as a new in vitro tool to study the molecular bases of FENIB.
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title_short	Glycosylation Tunes Neuroserpin Physiological and Pathological Properties
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Specific point mutations are responsible for its accumulation in the endoplasmic reticulum of neurons that leads to a pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers in vivo, while non-glycosylated NS causes aberrant polymer accumulation in cell models. To date, all in vitro studies have been conducted on bacterially expressed NS, de facto neglecting the role of glycosylation in the biochemical properties of NS. Here, we report the expression and purification of human glycosylated NS (gNS) using a novel eukaryotic expression system, LEXSY. Our results confirm the correct N-glycosylation of wild-type gNS. The fold and stability of gNS are not altered compared to bacterially expressed NS, as demonstrated by the circular dichroism and intrinsic tryptophan fluorescence assays. Intriguingly, gNS displays a remarkably reduced polymerisation propensity compared to non-glycosylated NS, in keeping with what was previously observed for wild-type NS in vivo and in cell models. 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Glycosylation Tunes Neuroserpin Physiological and Pathological Properties

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