A fluorescence resonance energy transfer sensor based on maltose binding protein

Igor L Medintz; Ellen R Goldman; Michael E Lassman; J Matthew Mauro

doi:10.1021/bc020062+

A fluorescence resonance energy transfer sensor based on maltose binding protein

Bioconjug Chem. 2003 Sep-Oct;14(5):909-18. doi: 10.1021/bc020062+.

Authors

Igor L Medintz¹, Ellen R Goldman, Michael E Lassman, J Matthew Mauro

Affiliation

¹ Center for Bio/Molecular Science and Engineering, Code 6900, US Naval Research Laboratory, Washington DC 20375, USA. Imedintz@cbmse.nrl.navy.mil

PMID: 13129393
DOI: 10.1021/bc020062+

Abstract

A fluorescence resonance energy-transfer (FRET) sensing system for maltose based on E. coli maltose binding protein (MBP) is demonstrated. The FRET donor portion of the sensing system consists of MBP modified with long wavelength-excitable cyanine dyes (Cy3 or Cy3.5). The novel acceptor portion of the sensor consists of beta-cyclodextrin (beta-CD) modified with either the cyanine dye Cy5 or the dark quencher QSY9. Binding of the modified beta-CD to dye-conjugated MBP results in assembly of the FRET complex. Added maltose displaces the beta-CD-dye adduct and disrupts the FRET complex, resulting in a direct change in fluorescence of the donor moiety. In the use of these FRET pairs, MBP dissociation values for maltose were estimated (0.14-2.90 microM). Maltose limits of detection were in the 50-100 nm range.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Biosensing Techniques / methods*
Carrier Proteins / analysis*
Carrier Proteins / chemical synthesis*
Fluorescence Resonance Energy Transfer / methods*
Maltose-Binding Proteins

Substances

Carrier Proteins
Maltose-Binding Proteins