Identification of glycosylation sites in the SU component of the Avian Sarcoma/Leukosis virus Envelope Glycoprotein (Subgroup A) by mass spectrometry

Mamuka Kvaratskhelia, Patrick K. Clark, Sonja Hess, Deborah C. Melder, Mark J Federspiel, Stephen H. Hughes

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

We used enzymatic digestion and mass spectrometry to identify the sites of glycosylation on the SU component of the Avian Sarcoma/Leukosis virus (ASLV) Envelope Glycoprotein (Subgroup A). The analysis was done with an SU(A)-rIgG fusion protein that binds the cognate receptor (Tva) specifically. PNGase F removed all the carbohydrate from the SU(A)-rIgG fusion. PNGase F is specific for N-linked carbohydrates; this shows that all the carbohydrate on SU(A) is N-linked. There are 10 modified aspargines in SU(A) (N17, N59, N80, N97, N117, N196, N230, N246, N254, and N330). All conform to the consensus site for N-linked glycosylation NXS/T. There is one potential glycosylation site (N236) that is not modified. Removing most of the carbohydrate from the mature SU(A)-rIgG by PNGase F treatment greatly reduces the ability of the protein to bind Tva, suggesting that carbohydrate may play a direct role in receptor binding.

Original languageEnglish (US)
Pages (from-to)171-181
Number of pages11
JournalVirology
Volume326
Issue number1
DOIs
StatePublished - Aug 15 2004

Fingerprint

Alpharetrovirus
Glycosylation
Mass Spectrometry
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Glycoproteins
Carbohydrates
Digestion
Proteins

Keywords

  • Avian
  • Envelope
  • Glycosylation
  • Retrovirus
  • Tva

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Identification of glycosylation sites in the SU component of the Avian Sarcoma/Leukosis virus Envelope Glycoprotein (Subgroup A) by mass spectrometry. / Kvaratskhelia, Mamuka; Clark, Patrick K.; Hess, Sonja; Melder, Deborah C.; Federspiel, Mark J; Hughes, Stephen H.

In: Virology, Vol. 326, No. 1, 15.08.2004, p. 171-181.

Research output: Contribution to journalArticle

Kvaratskhelia, Mamuka ; Clark, Patrick K. ; Hess, Sonja ; Melder, Deborah C. ; Federspiel, Mark J ; Hughes, Stephen H. / Identification of glycosylation sites in the SU component of the Avian Sarcoma/Leukosis virus Envelope Glycoprotein (Subgroup A) by mass spectrometry. In: Virology. 2004 ; Vol. 326, No. 1. pp. 171-181.
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