In vitro selection of GTP-binding proteins by block shuffling of estrogen-receptor fragments

Toru Tsuji, Michiko Onimaru, Nobuhide Doi, Etsuko Miyamoto-Sato, Hideaki Takashima, Hiroshi Yanagawa

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

To what extent has alternative splicing contributed to the evolution of protein-function diversity? We previously constructed a pool of block-deletion mutants of the human estrogen receptor α ligand binding domain by random multi-recombinant PCR. Here we performed iterative in vitro selection of GTP-binding proteins by using the library of mRNA-displayed proteins and GTP-affinity chromatography combined with quantitative real-time PCR. We obtained a novel GTP-binding protein with moderate affinity and substrate-specificity. The results of our in vitro simulation imply that alternative splicing may have contributed substantially to the diversification of protein function during evolution.

Original languageEnglish
Pages (from-to)689-693
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume390
Issue number3
DOIs
Publication statusPublished - 2009 Dec 18

Keywords

  • Alternative splicing
  • Combinatorial protein library
  • In vitro selection
  • Protein evolution
  • Synthetic biology
  • mRNA display

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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