The influence of N-glycosylation and C-terminal sequence on secretion of HBV large surface antigen from S. cerevisiae

Jin Seung Park, Hyuk Seong Seo, Jung Sun Yum, Hong Mo Moon, Jeewon Lee

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

In Saccharomyces cerevisiae, we synthesized and secreted L-HBVsAg (named as pre-S(Met1 to Asn174)::S(Met175 to Ile400)) and three mutants, i.e., pre-S ○○::S (Asn15Gln and Asn123Gln), pre-S ○○:: S (Asn15Gln, Asn123Gln, and Asn320Gln), and pre-S ○○::S ○○ (Asn15Gln, Asn123Gln, Asn233Gln, and Asn320Gln). All of the secreted pre-S::S was N-glycosylated, i.e., hyper-mannosylated. In the secretion of pre-S ○○::S and pre-S ○○::S , besides the hyper-mannosylated form, another immunoreactive protein with much lower molecular mass was observed, which seems to be unglycosylated form of pre-S ○○::S and pre-S ○○::S . Only a part of the secreted pre-S ○○::S or pre-S ○○::S molecules was N-glycosylated, and the site for the partial N-glycosylation seems to be Asn233 in S-antigen region. Compared to the N-glycosylated pre-S ○○::S and pre-S ○○::S , pre-S ○○::S ○○ (non-N-glycosylated mutant) was secreted with lower secretion efficiency but showed apparent immunoreactivity to anti-S antigen monoclonal Ab. Interestingly, unlike pre-S ○○::S ○○ with authentic C-terminus, the recombinant pre-S ○○::S ○○ with C-terminal myc or poly-histidine tag (pre-S ○○::S ○○::tag) was almost all aggregated into insoluble proteins in the intracellular region. Conclusively, the C-terminal sequence and glycosylation in S-antigen region seem to be of crucial importance in determining the secretion efficiency of L-HBVsAg in S. cerevisiae.

Original languageEnglish
Pages (from-to)250-255
Number of pages6
JournalBiotechnology and Bioengineering
Volume92
Issue number2
DOIs
Publication statusPublished - 2005 Oct 20

Fingerprint

Glycosylation
Surface Antigens
Antigens
Saccharomyces cerevisiae
Yeast
Proteins
Molecular mass
Molecules

Keywords

  • C-terminus of S antigen
  • L-HBVsAg
  • N-glycosylation
  • Secretion efficiency

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

Cite this

The influence of N-glycosylation and C-terminal sequence on secretion of HBV large surface antigen from S. cerevisiae. / Park, Jin Seung; Seo, Hyuk Seong; Yum, Jung Sun; Moon, Hong Mo; Lee, Jeewon.

In: Biotechnology and Bioengineering, Vol. 92, No. 2, 20.10.2005, p. 250-255.

Research output: Contribution to journalArticle

Park, Jin Seung ; Seo, Hyuk Seong ; Yum, Jung Sun ; Moon, Hong Mo ; Lee, Jeewon. / The influence of N-glycosylation and C-terminal sequence on secretion of HBV large surface antigen from S. cerevisiae. In: Biotechnology and Bioengineering. 2005 ; Vol. 92, No. 2. pp. 250-255.
@article{f44dcb7f10904761ae5b0eee3078b89a,
title = "The influence of N-glycosylation and C-terminal sequence on secretion of HBV large surface antigen from S. cerevisiae",
abstract = "In Saccharomyces cerevisiae, we synthesized and secreted L-HBVsAg (named as pre-S(Met1 to Asn174)::S(Met175 to Ile400)) and three mutants, i.e., pre-S ○○::S (Asn15Gln and Asn123Gln), pre-S ○○:: S ○ (Asn15Gln, Asn123Gln, and Asn320Gln), and pre-S ○○::S ○○ (Asn15Gln, Asn123Gln, Asn233Gln, and Asn320Gln). All of the secreted pre-S::S was N-glycosylated, i.e., hyper-mannosylated. In the secretion of pre-S ○○::S and pre-S ○○::S ○, besides the hyper-mannosylated form, another immunoreactive protein with much lower molecular mass was observed, which seems to be unglycosylated form of pre-S ○○::S and pre-S ○○::S ○. Only a part of the secreted pre-S ○○::S or pre-S ○○::S ○ molecules was N-glycosylated, and the site for the partial N-glycosylation seems to be Asn233 in S-antigen region. Compared to the N-glycosylated pre-S ○○::S and pre-S ○○::S ○, pre-S ○○::S ○○ (non-N-glycosylated mutant) was secreted with lower secretion efficiency but showed apparent immunoreactivity to anti-S antigen monoclonal Ab. Interestingly, unlike pre-S ○○::S ○○ with authentic C-terminus, the recombinant pre-S ○○::S ○○ with C-terminal myc or poly-histidine tag (pre-S ○○::S ○○::tag) was almost all aggregated into insoluble proteins in the intracellular region. Conclusively, the C-terminal sequence and glycosylation in S-antigen region seem to be of crucial importance in determining the secretion efficiency of L-HBVsAg in S. cerevisiae.",
keywords = "C-terminus of S antigen, L-HBVsAg, N-glycosylation, Secretion efficiency",
author = "Park, {Jin Seung} and Seo, {Hyuk Seong} and Yum, {Jung Sun} and Moon, {Hong Mo} and Jeewon Lee",
year = "2005",
month = "10",
day = "20",
doi = "10.1002/bit.20640",
language = "English",
volume = "92",
pages = "250--255",
journal = "Biotechnology and Bioengineering",
issn = "0006-3592",
publisher = "Wiley-VCH Verlag",
number = "2",

}

TY - JOUR

T1 - The influence of N-glycosylation and C-terminal sequence on secretion of HBV large surface antigen from S. cerevisiae

AU - Park, Jin Seung

AU - Seo, Hyuk Seong

AU - Yum, Jung Sun

AU - Moon, Hong Mo

AU - Lee, Jeewon

PY - 2005/10/20

Y1 - 2005/10/20

N2 - In Saccharomyces cerevisiae, we synthesized and secreted L-HBVsAg (named as pre-S(Met1 to Asn174)::S(Met175 to Ile400)) and three mutants, i.e., pre-S ○○::S (Asn15Gln and Asn123Gln), pre-S ○○:: S ○ (Asn15Gln, Asn123Gln, and Asn320Gln), and pre-S ○○::S ○○ (Asn15Gln, Asn123Gln, Asn233Gln, and Asn320Gln). All of the secreted pre-S::S was N-glycosylated, i.e., hyper-mannosylated. In the secretion of pre-S ○○::S and pre-S ○○::S ○, besides the hyper-mannosylated form, another immunoreactive protein with much lower molecular mass was observed, which seems to be unglycosylated form of pre-S ○○::S and pre-S ○○::S ○. Only a part of the secreted pre-S ○○::S or pre-S ○○::S ○ molecules was N-glycosylated, and the site for the partial N-glycosylation seems to be Asn233 in S-antigen region. Compared to the N-glycosylated pre-S ○○::S and pre-S ○○::S ○, pre-S ○○::S ○○ (non-N-glycosylated mutant) was secreted with lower secretion efficiency but showed apparent immunoreactivity to anti-S antigen monoclonal Ab. Interestingly, unlike pre-S ○○::S ○○ with authentic C-terminus, the recombinant pre-S ○○::S ○○ with C-terminal myc or poly-histidine tag (pre-S ○○::S ○○::tag) was almost all aggregated into insoluble proteins in the intracellular region. Conclusively, the C-terminal sequence and glycosylation in S-antigen region seem to be of crucial importance in determining the secretion efficiency of L-HBVsAg in S. cerevisiae.

AB - In Saccharomyces cerevisiae, we synthesized and secreted L-HBVsAg (named as pre-S(Met1 to Asn174)::S(Met175 to Ile400)) and three mutants, i.e., pre-S ○○::S (Asn15Gln and Asn123Gln), pre-S ○○:: S ○ (Asn15Gln, Asn123Gln, and Asn320Gln), and pre-S ○○::S ○○ (Asn15Gln, Asn123Gln, Asn233Gln, and Asn320Gln). All of the secreted pre-S::S was N-glycosylated, i.e., hyper-mannosylated. In the secretion of pre-S ○○::S and pre-S ○○::S ○, besides the hyper-mannosylated form, another immunoreactive protein with much lower molecular mass was observed, which seems to be unglycosylated form of pre-S ○○::S and pre-S ○○::S ○. Only a part of the secreted pre-S ○○::S or pre-S ○○::S ○ molecules was N-glycosylated, and the site for the partial N-glycosylation seems to be Asn233 in S-antigen region. Compared to the N-glycosylated pre-S ○○::S and pre-S ○○::S ○, pre-S ○○::S ○○ (non-N-glycosylated mutant) was secreted with lower secretion efficiency but showed apparent immunoreactivity to anti-S antigen monoclonal Ab. Interestingly, unlike pre-S ○○::S ○○ with authentic C-terminus, the recombinant pre-S ○○::S ○○ with C-terminal myc or poly-histidine tag (pre-S ○○::S ○○::tag) was almost all aggregated into insoluble proteins in the intracellular region. Conclusively, the C-terminal sequence and glycosylation in S-antigen region seem to be of crucial importance in determining the secretion efficiency of L-HBVsAg in S. cerevisiae.

KW - C-terminus of S antigen

KW - L-HBVsAg

KW - N-glycosylation

KW - Secretion efficiency

UR - http://www.scopus.com/inward/record.url?scp=27744507906&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=27744507906&partnerID=8YFLogxK

U2 - 10.1002/bit.20640

DO - 10.1002/bit.20640

M3 - Article

C2 - 16116655

AN - SCOPUS:27744507906

VL - 92

SP - 250

EP - 255

JO - Biotechnology and Bioengineering

JF - Biotechnology and Bioengineering

SN - 0006-3592

IS - 2

ER -