Chaperone-assisted folding of a single-chain antibody in a reconstituted translation system

Bei Wen Ying, Hideki Taguchi, Hiroshi Ueda, Takuya Ueda

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

A protein-synthesizing system based on a minimal set of purified components was used to investigate the roles molecular chaperones play in the folding of newly synthesized polypeptides. After we ascertained that this system lacks intrinsic chaperones, the effect of adding chaperones in a co-translational or post-translational manner was directly evaluated. An aggregation-prone single-chain antibody was used as the model nascent chain. The participation of the trigger factor or the DnaK system during translation efficiently increased the level of functional protein that was generated. In addition, both systems also acted as chaperones after translation had been stopped. In contrast, the GroEL/ES system showed little or no co- or post-translational assistance in folding.

Original languageEnglish
Pages (from-to)1359-1364
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume320
Issue number4
DOIs
Publication statusPublished - 2004 Aug 6
Externally publishedYes

Fingerprint

Single-Chain Antibodies
Molecular Chaperones
Proteins
Agglomeration
Peptides

Keywords

  • Cell-free translation
  • Co-translational
  • Molecular chaperone
  • Post-translational
  • Protein folding
  • scFv

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Chaperone-assisted folding of a single-chain antibody in a reconstituted translation system. / Ying, Bei Wen; Taguchi, Hideki; Ueda, Hiroshi; Ueda, Takuya.

In: Biochemical and Biophysical Research Communications, Vol. 320, No. 4, 06.08.2004, p. 1359-1364.

Research output: Contribution to journalArticle

Ying, Bei Wen ; Taguchi, Hideki ; Ueda, Hiroshi ; Ueda, Takuya. / Chaperone-assisted folding of a single-chain antibody in a reconstituted translation system. In: Biochemical and Biophysical Research Communications. 2004 ; Vol. 320, No. 4. pp. 1359-1364.
@article{5477e379ff2a41eaa25cc38e7078490e,
title = "Chaperone-assisted folding of a single-chain antibody in a reconstituted translation system",
abstract = "A protein-synthesizing system based on a minimal set of purified components was used to investigate the roles molecular chaperones play in the folding of newly synthesized polypeptides. After we ascertained that this system lacks intrinsic chaperones, the effect of adding chaperones in a co-translational or post-translational manner was directly evaluated. An aggregation-prone single-chain antibody was used as the model nascent chain. The participation of the trigger factor or the DnaK system during translation efficiently increased the level of functional protein that was generated. In addition, both systems also acted as chaperones after translation had been stopped. In contrast, the GroEL/ES system showed little or no co- or post-translational assistance in folding.",
keywords = "Cell-free translation, Co-translational, Molecular chaperone, Post-translational, Protein folding, scFv",
author = "Ying, {Bei Wen} and Hideki Taguchi and Hiroshi Ueda and Takuya Ueda",
year = "2004",
month = "8",
day = "6",
doi = "10.1016/j.bbrc.2004.06.095",
language = "English",
volume = "320",
pages = "1359--1364",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "4",

}

TY - JOUR

T1 - Chaperone-assisted folding of a single-chain antibody in a reconstituted translation system

AU - Ying, Bei Wen

AU - Taguchi, Hideki

AU - Ueda, Hiroshi

AU - Ueda, Takuya

PY - 2004/8/6

Y1 - 2004/8/6

N2 - A protein-synthesizing system based on a minimal set of purified components was used to investigate the roles molecular chaperones play in the folding of newly synthesized polypeptides. After we ascertained that this system lacks intrinsic chaperones, the effect of adding chaperones in a co-translational or post-translational manner was directly evaluated. An aggregation-prone single-chain antibody was used as the model nascent chain. The participation of the trigger factor or the DnaK system during translation efficiently increased the level of functional protein that was generated. In addition, both systems also acted as chaperones after translation had been stopped. In contrast, the GroEL/ES system showed little or no co- or post-translational assistance in folding.

AB - A protein-synthesizing system based on a minimal set of purified components was used to investigate the roles molecular chaperones play in the folding of newly synthesized polypeptides. After we ascertained that this system lacks intrinsic chaperones, the effect of adding chaperones in a co-translational or post-translational manner was directly evaluated. An aggregation-prone single-chain antibody was used as the model nascent chain. The participation of the trigger factor or the DnaK system during translation efficiently increased the level of functional protein that was generated. In addition, both systems also acted as chaperones after translation had been stopped. In contrast, the GroEL/ES system showed little or no co- or post-translational assistance in folding.

KW - Cell-free translation

KW - Co-translational

KW - Molecular chaperone

KW - Post-translational

KW - Protein folding

KW - scFv

UR - http://www.scopus.com/inward/record.url?scp=3142771251&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3142771251&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2004.06.095

DO - 10.1016/j.bbrc.2004.06.095

M3 - Article

C2 - 15303282

AN - SCOPUS:3142771251

VL - 320

SP - 1359

EP - 1364

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 4

ER -