Photophysics of ANS. I. Protein–ANS complexes: Intestinal fatty acid binding protein and single-trp mutants

Klimtchuk, Elena; Venyaminov, Sergei Yu.; Kurian, Elizabeth; Wessels, William S.; Kirk, William R.; Prendergast, Franklyn G.

doi:10.1016/j.bpc.2006.07.016

Cited by 14 publications

(11 citation statements)

References 21 publications

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“…This correction is not always needed depending on the affinities of ANS and fatty acids. In the case of Cq-FABP, the apparent K d,ANS value was 30 µM, significantly higher than the value previously found for other FABPs (in the low micromolar range for I-FABP and L-FABP) [41]–[43], and thus required such a correction. In contrast to the “intensity” procedure, the fluorescence of free ANS is negligible when monitoring the fluorescence emission of Trp in the “FRET” procedure.…”

Section: Discussioncontrasting

confidence: 62%

“…4 & 5). It is important to note that, in the absence of any structural data for Cq-FABP, it is difficult to certify if the observed quenching of Trp can be simply explained by the energy transfer theory (based on a weak dipole-dipole interaction) or by another mechanism involving exciton coupling as previously suggested for intestinal FABP (I-FABP) [41].…”

Section: Discussionmentioning

confidence: 98%

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Comparative Study of the Fatty Acid Binding Process of a New FABP from Cherax quadricarinatus by Fluorescence Intensity, Lifetime and Anisotropy

Henry

Wang

et al. 2012

PLoS ONE

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Fatty acid-binding proteins (FABPs) are small cytosolic proteins, largely distributed in invertebrates and vertebrates, which accomplish uptake and intracellular transport of hydrophobic ligands such as fatty acids. Although long chain fatty acids play multiple crucial roles in cellular functions (structural, energy metabolism, regulation of gene expression), the precise functions of FABPs, especially those of invertebrate species, remain elusive. Here, we have identified and characterized a novel FABP family member, Cq-FABP, from the hepatopancreas of red claw crayfish Cherax quadricarinatus. We report the characterization of fatty acid-binding affinity of Cq-FABP by four different competitive fluorescence-based assays. In the two first approaches, the fluorescent probe 8-Anilino-1-naphthalenesulfonate (ANS), a binder of internal cavities of protein, was used either by directly monitoring its fluorescence emission or by monitoring the fluorescence resonance energy transfer occurring between the single tryptophan residue of Cq-FABP and ANS. The third and the fourth approaches were based on the measurement of the fluorescence emission intensity of the naturally fluorescent cis-parinaric acid probe or the steady-state fluorescence anisotropy measurements of a fluorescently labeled fatty acid (BODIPY-C16), respectively. The four methodologies displayed consistent equilibrium constants for a given fatty acid but were not equivalent in terms of analysis. Indeed, the two first methods were complicated by the existence of non specific binding modes of ANS while BODIPY-C16 and cis-parinaric acid specifically targeted the fatty acid binding site. We found a relationship between the affinity and the length of the carbon chain, with the highest affinity obtained for the shortest fatty acid, suggesting that steric effects primarily influence the interaction of fatty acids in the binding cavity of Cq-FABP. Moreover, our results show that the binding affinities of several fatty acids closely parallel their prevalences in the hepatopancreas of C. quadricarinatus as measured under specific diet conditions.

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Section: Discussioncontrasting

confidence: 62%

Section: Discussionmentioning

confidence: 98%

Comparative Study of the Fatty Acid Binding Process of a New FABP from Cherax quadricarinatus by Fluorescence Intensity, Lifetime and Anisotropy

Henry

Wang

et al. 2012

PLoS ONE

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“…As quantum mechanic calculations suggested, 1-substituted naphthalenes exhibit mainly a-type activity, i.e., the S 0 → S 1 transitions of 1-substituted naphthalenes is polarized along the long in-plane axis [24]. Therefore, the transitional direction of ANS molecule is along its long axis [25,26] (as shown in Scheme 1).…”

Section: Fluorescence Polarizationmentioning

confidence: 95%

Preparation of an anion dye intercalated into layered double hydroxides and its controllable luminescence properties

Sun

Liu

Shi

et al. 2010

Chemical Engineering Journal

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“…The decays of the protein in absence and presence of ligand (Figure 5) were fitted with triexponential functions (see Experimental section for details) and the computed lifetimes, ascribed to different conformers of the Trp residues [24], are listed in Table 2. It must be stressed that the τ 1 value is below the resolution of our device and could not accurately be determined.…”

Section: Resultsmentioning

confidence: 99%

2-(2-Hydroxy-5-nitrobenzylidene)-1,3-indanedione versus Fluorescein Isothiocyanate in Interaction with Anti-hFABP Immunoglobulin G1: Fluorescence Quenching, Secondary Structure Alteration and Binding Sites Localization

Stan

Mihailescu

Savin

et al. 2013

IJMS

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The first step in determining whether a fluorescent dye can be used for antibody labeling consists in collecting data on its physical interaction with the latter. In the present study, the interaction between the 2-(2-hydroxy-5-nitrobenzylidene)-1,3-indanedione (HNBID) dye and the IgG1 monoclonal mouse antibody anti-human heart fatty acid binding protein (anti-hFABP) has been investigated by fluorescence and circular dichroism spectroscopies and complementary structural results were obtained by molecular modeling. We have determined the parameters characterizing this interaction, namely the quenching and binding constants, classes of binding sites, and excited state lifetimes, and we have predicted the localization of HNBID within the Fc region of anti-hFABP. The key glycosidic and amino acid residues in anti-hFABP interacting with HNBID have also been identified. A similar systematic study was undertaken for the well-known fluorescein isothiocyanate fluorophore, for comparison purposes. Our results recommend HNBID as a valuable alternative to fluorescein isothiocyanate for use as a fluorescent probe for IgG1 antibodies.

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Photophysics of ANS. I. Protein–ANS complexes: Intestinal fatty acid binding protein and single-trp mutants

Cited by 14 publications

References 21 publications

Comparative Study of the Fatty Acid Binding Process of a New FABP from Cherax quadricarinatus by Fluorescence Intensity, Lifetime and Anisotropy

Comparative Study of the Fatty Acid Binding Process of a New FABP from Cherax quadricarinatus by Fluorescence Intensity, Lifetime and Anisotropy

Preparation of an anion dye intercalated into layered double hydroxides and its controllable luminescence properties

2-(2-Hydroxy-5-nitrobenzylidene)-1,3-indanedione versus Fluorescein Isothiocyanate in Interaction with Anti-hFABP Immunoglobulin G1: Fluorescence Quenching, Secondary Structure Alteration and Binding Sites Localization

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