Characterization of random-sequence proteins displayed on the surface of Escherichia coli RNase HI

Nobuhide Doi, Tetsuya Yomo, Mitsuhiro Itaya, Hiroshi Yanagawa

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

In a previous study, random-sequence proteins of 120-130 amino acid residues mere inserted into the surface loop region of the enzyme, Escherichia coli RNase HI [Doi et al. (1997) FEBS Lett. 102, 177-180]. Here we established that the RNase H activity of the insertion mutants is correlated with their secondary structure contents evaluated by circular dichroism measurement at 222 nm. The random-sequence insert of a mutant enzyme possessing relatively high RNase H activity was detached from the RNase HI scaffold, and its characterization indicated that the random-sequence protein maintains its secondary structure after separation from the scaffold. Thus, the structural features of random-sequence proteins were suggested to be monitored by measuring the activity of the scaffold enzyme into which these proteins have been inserted.

Original languageEnglish
Pages (from-to)51-54
Number of pages4
JournalFEBS Letters
Volume427
Issue number1
DOIs
Publication statusPublished - 1998 May 1
Externally publishedYes

Fingerprint

Escherichia coli
Scaffolds
Ribonuclease H
Proteins
Enzymes
Circular Dichroism
Amino Acids
ribonuclease HI

Keywords

  • Directed evolution
  • Evolutionary engineering
  • Insertional mutagenesis
  • Protein folding
  • Protein scaffold

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Characterization of random-sequence proteins displayed on the surface of Escherichia coli RNase HI. / Doi, Nobuhide; Yomo, Tetsuya; Itaya, Mitsuhiro; Yanagawa, Hiroshi.

In: FEBS Letters, Vol. 427, No. 1, 01.05.1998, p. 51-54.

Research output: Contribution to journalArticle

Doi, Nobuhide ; Yomo, Tetsuya ; Itaya, Mitsuhiro ; Yanagawa, Hiroshi. / Characterization of random-sequence proteins displayed on the surface of Escherichia coli RNase HI. In: FEBS Letters. 1998 ; Vol. 427, No. 1. pp. 51-54.
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