Photoacoustic analysis of proteins: Volumetric signals and fluorescence quantum yields

Elizabeth Kurian, Franklyn G. Prendergast, Jeanne Rudzki Small

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

A series of proteins has been examined using time-resolved, pulsed- laser volumetric photoacoustic spectroscopy. Photoacoustic waveforms were collected to measure heat release for calculation of fluorescence quantum yields, and to explore the possibility of photoinduced nonthermal volume changes occurring in these protein samples. The proteins studied were the green fluorescent protein (GFP); intestinal fatty acid binding protein (IFABP), and adipocyte lipid-binding protein (ALBP), each labeled noncovalently with 1-anilinonaphthalene-8-sulfonate (1,8-ANS) and covalently with 6-acryloyl-2-(dimethylamino)naphthalene (acrylodan); and acrylodan- labeled IFABP and ALBP with added oleic acid. Of this group of proteins, only the ALBP labeled with 1,8-ANS showed significant nonthermal volume changes at the β = 0 temperature (~3.8°C) for the buffer used (10 mM Tris-HCl, pH 7.5) (β is the thermal cubic volumetric expansion coefficient). For all of the proteins except for acrylodan-labeled IFABP, the fluorescence quantum yields calculated assuming simple energy conservation were anomalously high, i.e., the apparent heat signals were lower than those predicted from independent fluorescence measurements. The consistent anomalies suggest that the low photoacoustic signals may be characteristic of fluorophores buried in proteins, and that photoacoustic signals derive in part from the microenvironment of the absorbing chromophore.

Original languageEnglish (US)
Pages (from-to)466-476
Number of pages11
JournalBiophysical Journal
Volume73
Issue number1
DOIs
StatePublished - Jul 1997

ASJC Scopus subject areas

  • Biophysics

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