Single-Molecule Spectroscopy Selectively Probes Donor and Acceptor Chromophores in the Phycobiliprotein Allophycocyanin

Davey Loos, Mircea Cotlet*, Frans De Schryver, Satoshi Habuchi, Johan Hofkens

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

46 Scopus citations


We report on single-molecule fluorescence measurements performed on the phycobiliprotein allophycocyanin (APC). Our data support the presence of a unidirectional Förster-type energy transfer process involving spectrally different chromophores, α84 (donor) and β84 (acceptor), as well as of energy hopping amongst β84 chromophores. Single-molecule fluorescence spectra recorded from individual immobilized APC proteins indicate the presence of a red-emitting chromophore with emission peaking at 660 nm, which we connect with β84, and a species with the emission peak blue shifted at 630 nm, which we attribute to α84. Polarization data from single APC trimers point to the presence of three consecutive red emitters, suggesting energy hopping amongst β84 chromophores. Based on the single-molecule fluorescence spectra and assuming that emission at the ensemble level in solution comes mainly from the acceptor chromophore, we were able to resolve the individual absorption and emission spectra of the α84 and β84 chromophores in APC.

Original languageEnglish (US)
Pages (from-to)2598-2608
Number of pages11
JournalBiophysical Journal
Issue number4
StatePublished - Oct 2004

Bibliographical note

Funding Information:
D.L. thanks the Instituut voor Innovatie door Wetenschap en Technologie and Katholieke Universiteit Leuven (KUL) for financial support. M.C. thanks Onderzoeksfonds of KUL for a postdoctoral fellowship. S.H. thanks the Japan Society for the Promotion of Science for a postdoctoral fellowship. The federal science policy through IAP/V/03 and the KUL through GOA 01/2 are thanked.

Publisher Copyright:
© 2004 The Biophysical Society

ASJC Scopus subject areas

  • Biophysics


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