Full Paper

A Genetically Encoded Pore for the Stochastic Detection of a Protein Kinase

  1. Stephen Cheley Dr.2,3,†,
  2. Hongzhi Xie Dr.2,4,†,
  3. Hagan Bayley Prof. Dr.1

Article first published online: 27 OCT 2006

DOI: 10.1002/cbic.200600274

Issue

ChemBioChem

ChemBioChem

Volume 7, Issue 12, pages 1923–1927, December 4, 2006

Additional Information

How to Cite

Cheley, S., Xie, H. and Bayley, H. (2006), A Genetically Encoded Pore for the Stochastic Detection of a Protein Kinase. ChemBioChem, 7: 1923–1927. doi: 10.1002/cbic.200600274

Author Information

  1. 1

    Department of Chemistry, University of Oxford, Mansfield Road, Oxford, OX1 3TA, England, UK, Fax: (+44) 1865-275708

  2. 2

    Department of Molecular and Cellular Medicine, The Texas A&M University System Health Science Center, College Station, TX 77843-1114, USA

  3. 3

    Present address: Department of Chemistry, Michigan State University, East Lansing, MI 48824, USA

  4. 4

    Present address: Los Alamos National Laboratory, Chemistry Division, C-PCS Group, P. O. Box 1663, MS J 567, Los Alamos, NM 87545, USA

  1. †

    These authors contributed equally to this work.

Publication History

  1. Issue published online: 27 NOV 2006
  2. Article first published online: 27 OCT 2006
  3. Manuscript Received: 6 JUL 2006

Funded by

  • DARPA
  • NASA
  • NIH
  • ONR

SEARCH

SEARCH BY CITATION

ARTICLE TOOLS

View Full Article with Supporting Information (HTML) Get PDF (364K)

Keywords:

  • kinases;
  • pores;
  • protein engineering;
  • single-molecule detection;
  • stochastic sensing

Graphical Abstract

Thumbnail image of graphical abstract

The genetic way. Protein analytes can be detected by stochastic sensing in which the modulation of a current flowing through a single protein pore is observed. In previous work, proteins have been detected with pores to which ligands were appended by targeted chemical modification. We now report the first genetically encoded stochastic sensor element for detecting a protein: an α-hemolysin pore containing a single copy of a protein kinase inhibitor sequence. This development should facilitate the rapid screening of kinase inhibitors.

Abstract

Stochastic sensing is an emerging approach for the detection of a wide variety of analytes at the level of individual molecules. Detection is accomplished by observing the modulation of the current that flows through a single protein pore that has been engineered to bind an analyte of interest. Previously, protein analytes have been detected by using pores to which ligands have been appended at specific sites by targeted chemical modification. Here, we report the first genetically encoded stochastic sensor element for detecting a protein. A protein kinase inhibitor peptide sequence was incorporated into the α-hemolysin polypeptide, which was used to form a heteroheptameric pore containing a single copy of the inhibitor sequence. With this pore, the successful detection of the catalytic subunit of protein kinase A was demonstrated. This development should greatly facilitate the detection of active kinase subunits by stochastic sensing and the rapid screening of kinase inhibitors by an approach that yields kinetic information.

View Full Article with Supporting Information (HTML) Get PDF (364K)