RIEDL tag: A novel pentapeptide tagging system for transmembrane protein purification

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

Affinity tag systems are an essential tool in biochemistry, biophysics, and molecular biology. Although several different tag systems have been developed, the epitope tag system, composed of a polypeptide “tag” and an anti-tag antibody, is especially useful for protein purification. However, almost all tag sequences, such as the FLAG tag, are added to the N- or C-termini of target proteins, as tags inserted in loops tend to disrupt the functional structure of multi-pass transmembrane proteins. In this study, we developed a novel “RIEDL tag system,” which is composed of a peptide with only five amino acids (RIEDL) and an anti-RIEDL monoclonal antibody (mAb), LpMab-7. To investigate whether the RIEDL tag system is applicable for protein purification, we conducted the purification of two kinds of RIEDL-tagged proteins using affinity column chromatography: whale podoplanin (wPDPN) with an N-terminal RIEDL tag (RIEDL-wPDPN) and human CD20 with an internal RIEDL tag insertion (CD20-169RIEDL170). Using an LpMab-7-Sepharose column, RIEDL-wPDPN and CD20-169RIEDL170 were efficiently purified in one-step purification procedures, and were strongly detected by LpMab-7 using Western blot and flow cytometry. These results show that the RIEDL tag system can be useful for the detection and one-step purification of membrane proteins when inserted at either the N-terminus or inserted in an internal loop structure of multi-pass transmembrane proteins.

Original languageEnglish
Article number100780
JournalBiochemistry and Biophysics Reports
Volume23
DOIs
Publication statusPublished - 2020 Sep

Keywords

  • CD20
  • Monoclonal antibody
  • Peptide tag
  • Podoplanin
  • Protein purification
  • RIEDL tag

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

Fingerprint

Dive into the research topics of 'RIEDL tag: A novel pentapeptide tagging system for transmembrane protein purification'. Together they form a unique fingerprint.

Cite this