Gp120 on HIV-1 virions lacks O-linked carbohydrate

Elizabeth Stansell, Maria Panico, Kevin Canis, Poh Choo Pang, Laura Bouché, Daniel Binet, Michael John O'Connor, Elena Chertova, Julian Bess, Jeffrey D. Lifson, Stuart M. Haslam, Howard R. Morris, Ronald Charles Desrosiers, Anne Dell

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

As HIV-1-encoded envelope protein traverses the secretory pathway, it may be modified with N- and O-linked carbohydrate. When the gp120s of HIV-1 NL4-3, HIV-1 YU2, HIV-1 Bal, HIV-1 JRFL, and HIV-1 JRCSF were expressed as secreted proteins, the threonine at consensus position 499 was found to be O-glycosylated. For SIVmac239, the corresponding threonine was also glycosylated when gp120 was recombinantly expressed. Similarly-positioned, highly-conserved threonines in the influenza A virus H1N1 HA1 and H5N1 HA1 envelope proteins were also found to carry O-glycans when expressed as secreted proteins. In all cases, the threonines were modified predominantly with disialylated core 1 glycans, together with related core 1 and core 2 structures. Secreted HIV-1 gp140 was modified to a lesser extent with mainly monosialylated core 1 O-glycans, suggesting that the ectodomain of the gp41 transmembrane component may limit the accessibility of Thr499 to glycosyltransferases. In striking contrast to these findings, gp120 on purified virions of HIV-1 Bal and SIV CP-MAC lacked any detectable O-glycosylation of the C-terminal threonine. Our results indicate the absence of O-linked carbohydrates on Thr499 as it exists on the surface of virions and suggest caution in the interpretation of analyses of post-translational modifications that utilize recombinant forms of envelope protein.

Original languageEnglish (US)
Article numbere0124784
JournalPLoS One
Volume10
Issue number4
DOIs
StatePublished - Apr 27 2015

Fingerprint

HIV Envelope Protein gp120
Threonine
Human immunodeficiency virus 1
virion
Virion
HIV-1
Carbohydrates
carbohydrates
threonine
Polysaccharides
Proteins
polysaccharides
Glycosylation
Glycosyltransferases
proteins
Viruses
Secretory Pathway
Influenza A virus
glycosyltransferases
Post Translational Protein Processing

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Stansell, E., Panico, M., Canis, K., Pang, P. C., Bouché, L., Binet, D., ... Dell, A. (2015). Gp120 on HIV-1 virions lacks O-linked carbohydrate. PLoS One, 10(4), [e0124784]. https://doi.org/10.1371/journal.pone.0124784

Gp120 on HIV-1 virions lacks O-linked carbohydrate. / Stansell, Elizabeth; Panico, Maria; Canis, Kevin; Pang, Poh Choo; Bouché, Laura; Binet, Daniel; O'Connor, Michael John; Chertova, Elena; Bess, Julian; Lifson, Jeffrey D.; Haslam, Stuart M.; Morris, Howard R.; Desrosiers, Ronald Charles; Dell, Anne.

In: PLoS One, Vol. 10, No. 4, e0124784, 27.04.2015.

Research output: Contribution to journalArticle

Stansell, E, Panico, M, Canis, K, Pang, PC, Bouché, L, Binet, D, O'Connor, MJ, Chertova, E, Bess, J, Lifson, JD, Haslam, SM, Morris, HR, Desrosiers, RC & Dell, A 2015, 'Gp120 on HIV-1 virions lacks O-linked carbohydrate', PLoS One, vol. 10, no. 4, e0124784. https://doi.org/10.1371/journal.pone.0124784
Stansell E, Panico M, Canis K, Pang PC, Bouché L, Binet D et al. Gp120 on HIV-1 virions lacks O-linked carbohydrate. PLoS One. 2015 Apr 27;10(4). e0124784. https://doi.org/10.1371/journal.pone.0124784
Stansell, Elizabeth ; Panico, Maria ; Canis, Kevin ; Pang, Poh Choo ; Bouché, Laura ; Binet, Daniel ; O'Connor, Michael John ; Chertova, Elena ; Bess, Julian ; Lifson, Jeffrey D. ; Haslam, Stuart M. ; Morris, Howard R. ; Desrosiers, Ronald Charles ; Dell, Anne. / Gp120 on HIV-1 virions lacks O-linked carbohydrate. In: PLoS One. 2015 ; Vol. 10, No. 4.
@article{f8350f1b3f8849edb029e7adde193a4a,
title = "Gp120 on HIV-1 virions lacks O-linked carbohydrate",
abstract = "As HIV-1-encoded envelope protein traverses the secretory pathway, it may be modified with N- and O-linked carbohydrate. When the gp120s of HIV-1 NL4-3, HIV-1 YU2, HIV-1 Bal, HIV-1 JRFL, and HIV-1 JRCSF were expressed as secreted proteins, the threonine at consensus position 499 was found to be O-glycosylated. For SIVmac239, the corresponding threonine was also glycosylated when gp120 was recombinantly expressed. Similarly-positioned, highly-conserved threonines in the influenza A virus H1N1 HA1 and H5N1 HA1 envelope proteins were also found to carry O-glycans when expressed as secreted proteins. In all cases, the threonines were modified predominantly with disialylated core 1 glycans, together with related core 1 and core 2 structures. Secreted HIV-1 gp140 was modified to a lesser extent with mainly monosialylated core 1 O-glycans, suggesting that the ectodomain of the gp41 transmembrane component may limit the accessibility of Thr499 to glycosyltransferases. In striking contrast to these findings, gp120 on purified virions of HIV-1 Bal and SIV CP-MAC lacked any detectable O-glycosylation of the C-terminal threonine. Our results indicate the absence of O-linked carbohydrates on Thr499 as it exists on the surface of virions and suggest caution in the interpretation of analyses of post-translational modifications that utilize recombinant forms of envelope protein.",
author = "Elizabeth Stansell and Maria Panico and Kevin Canis and Pang, {Poh Choo} and Laura Bouch{\'e} and Daniel Binet and O'Connor, {Michael John} and Elena Chertova and Julian Bess and Lifson, {Jeffrey D.} and Haslam, {Stuart M.} and Morris, {Howard R.} and Desrosiers, {Ronald Charles} and Anne Dell",
year = "2015",
month = "4",
day = "27",
doi = "10.1371/journal.pone.0124784",
language = "English (US)",
volume = "10",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "4",

}

TY - JOUR

T1 - Gp120 on HIV-1 virions lacks O-linked carbohydrate

AU - Stansell, Elizabeth

AU - Panico, Maria

AU - Canis, Kevin

AU - Pang, Poh Choo

AU - Bouché, Laura

AU - Binet, Daniel

AU - O'Connor, Michael John

AU - Chertova, Elena

AU - Bess, Julian

AU - Lifson, Jeffrey D.

AU - Haslam, Stuart M.

AU - Morris, Howard R.

AU - Desrosiers, Ronald Charles

AU - Dell, Anne

PY - 2015/4/27

Y1 - 2015/4/27

N2 - As HIV-1-encoded envelope protein traverses the secretory pathway, it may be modified with N- and O-linked carbohydrate. When the gp120s of HIV-1 NL4-3, HIV-1 YU2, HIV-1 Bal, HIV-1 JRFL, and HIV-1 JRCSF were expressed as secreted proteins, the threonine at consensus position 499 was found to be O-glycosylated. For SIVmac239, the corresponding threonine was also glycosylated when gp120 was recombinantly expressed. Similarly-positioned, highly-conserved threonines in the influenza A virus H1N1 HA1 and H5N1 HA1 envelope proteins were also found to carry O-glycans when expressed as secreted proteins. In all cases, the threonines were modified predominantly with disialylated core 1 glycans, together with related core 1 and core 2 structures. Secreted HIV-1 gp140 was modified to a lesser extent with mainly monosialylated core 1 O-glycans, suggesting that the ectodomain of the gp41 transmembrane component may limit the accessibility of Thr499 to glycosyltransferases. In striking contrast to these findings, gp120 on purified virions of HIV-1 Bal and SIV CP-MAC lacked any detectable O-glycosylation of the C-terminal threonine. Our results indicate the absence of O-linked carbohydrates on Thr499 as it exists on the surface of virions and suggest caution in the interpretation of analyses of post-translational modifications that utilize recombinant forms of envelope protein.

AB - As HIV-1-encoded envelope protein traverses the secretory pathway, it may be modified with N- and O-linked carbohydrate. When the gp120s of HIV-1 NL4-3, HIV-1 YU2, HIV-1 Bal, HIV-1 JRFL, and HIV-1 JRCSF were expressed as secreted proteins, the threonine at consensus position 499 was found to be O-glycosylated. For SIVmac239, the corresponding threonine was also glycosylated when gp120 was recombinantly expressed. Similarly-positioned, highly-conserved threonines in the influenza A virus H1N1 HA1 and H5N1 HA1 envelope proteins were also found to carry O-glycans when expressed as secreted proteins. In all cases, the threonines were modified predominantly with disialylated core 1 glycans, together with related core 1 and core 2 structures. Secreted HIV-1 gp140 was modified to a lesser extent with mainly monosialylated core 1 O-glycans, suggesting that the ectodomain of the gp41 transmembrane component may limit the accessibility of Thr499 to glycosyltransferases. In striking contrast to these findings, gp120 on purified virions of HIV-1 Bal and SIV CP-MAC lacked any detectable O-glycosylation of the C-terminal threonine. Our results indicate the absence of O-linked carbohydrates on Thr499 as it exists on the surface of virions and suggest caution in the interpretation of analyses of post-translational modifications that utilize recombinant forms of envelope protein.

UR - http://www.scopus.com/inward/record.url?scp=84928609224&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84928609224&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0124784

DO - 10.1371/journal.pone.0124784

M3 - Article

C2 - 25915761

AN - SCOPUS:84928609224

VL - 10

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 4

M1 - e0124784

ER -