Renaturation and stabilization of the telomere-binding activity of Saccharomyces Cdc13(451-693)p by L-arginine

Yi Chien Lin, Jing Wen Shih, Chia Ling Hsu, Jing Jer Lin

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Production of recombinant proteins can be valuable in studying their biological functions. However, recombinant proteins expressed in Escherichia coli sometimes form undesirable insoluble aggregates. Solubilization and renaturation of these aggregates becomes a problem that one needs to solve. Here we used recombinant Cdc13(451-693)p as example to show the presence of L-arginine during renaturation greatly enhanced the renaturation efficiency. Cdc13p is the single-stranded telomere-binding protein of yeast Saccharomyces cerevisiae. The telomere-binding domain has been mapped within amino acids 451-693 of Cdc13p, Cdc13(451-693)p. Recombinant Cdc13(451-693)p was expressed in E. coli as insoluble protein aggregates. Purification of insoluble Cdc13(451-693)p was achieved by denaturing the protein with 6 M guanidine-HCl and followed by Ni-nitrilotriacetic acid agarose column chromatography. Renaturation of Cdc13(451-693)p to the active form was achieved by dialyzing denatured protein in the presence of L-arginine. Moreover, the presence of L-arginine was also helped in maintaining the telomere-binding activity of Cdc13(451-693)p. Taking together, L-arginine might have a general application in renaturation of insoluble aggregates.

Original languageEnglish
Pages (from-to)44-47
Number of pages4
JournalAnalytical Biochemistry
Volume294
Issue number1
DOIs
Publication statusPublished - Jul 1 2001
Externally publishedYes

Fingerprint

Saccharomyces
Telomere
Arginine
Stabilization
Recombinant Proteins
Yeast
Escherichia coli
Telomere-Binding Proteins
Nitrilotriacetic Acid
Agarose Chromatography
Column chromatography
Guanidine
Sepharose
Purification
Saccharomyces cerevisiae
Proteins
Yeasts
Amino Acids

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Renaturation and stabilization of the telomere-binding activity of Saccharomyces Cdc13(451-693)p by L-arginine. / Lin, Yi Chien; Shih, Jing Wen; Hsu, Chia Ling; Lin, Jing Jer.

In: Analytical Biochemistry, Vol. 294, No. 1, 01.07.2001, p. 44-47.

Research output: Contribution to journalArticle

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