Fluorescence-Line-Narrowed Spectra of Polycyclic Aromatic Carcinogen-DNA Adducts

Heisig, V.; Jeffrey, Alan M.; McGlade, Michael J.; Small, Gerald J.

doi:10.1126/science.6422551

Cited by 29 publications

(29 citation statements)

References 10 publications

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“…Recent attempts at identifying DNA-PAH adducts have led to the development of several techniques, such as enzyme-linked immunosorbent assay (ELISA) and Randerath procedure (8)(9)(10). A fluorescence-based approach which affords high selectivity and one that has been applied to intact DNA adducts is fluorescence line narrowing (FLN) spectrometry (11)(12)(13)(14)(15)(16)(17)(18)(19)(20). In this methodology the 0893-228x/88/2701-0060$01.50/0 analytes are dissolved in a glass-forming solvent which is cooled to cryogenic temperatures (typically 4-20 K).…”

Section: Introductionmentioning

confidence: 99%

“…II. Principles of Fluorescence Line Narrowing and Hole Burning Fluorescence line narrowing (11)(12)(13)(14)(15)(16)(17)(18)(19)(20) and spectral hole burning (21)(22)(23) are site excitation energy (isochromat) selective spectroscopies that can eliminate or significantly reduce the contribution of site inhomogeneous line broadening, rt, to spectral profiles. A site inhomogeneously broadened absorption profile is depicted in Figure la and is the convolution of a very large number of individual site absorptions possessing a homogeneous line width T. At liquid helium temperatures T < 0.1 cm'1, while for amorphous molecular hosts such as glasses or polymers, Ti ~300-500 cm'1.…”

Section: Introductionmentioning

confidence: 99%

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Fluorescence line narrowing-nonphotochemical hole burning spectrometry: femtomole detection and high selectivity for intact DNA-PAH adducts

Jankowiak¹,

Zamzow²,

Small³

et al. 1988

Chem. Res. Toxicol.

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A new fluorescence line narrowing (FLN) apparatus is described and evaluated through experiments on intact DNA-PAH (polycyclic aromatic hydrocarbon) and globin-PAH adducts, as well as polar PAH metabolites. A detection limit of approximately 3 modified bases in 10(8) for a DNA adduct formed with a diol-epoxide of benzo[a]pyrene (BPDE-DNA) is reported for 20 micrograms of DNA at a spectral resolution of approximately 8 cm-1. The methodology employed avoids or minimizes spectral degradation and loss of sensitivity due to photooxidation and nonphotochemical hole burning (NPHB). A new double selection technique that employs both FLN and NPHB is described and found to lead to a significant improvement in selectivity over that obtained with conventional FLN.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fluorescence line narrowing-nonphotochemical hole burning spectrometry: femtomole detection and high selectivity for intact DNA-PAH adducts

Jankowiak¹,

Zamzow²,

Small³

et al. 1988

Chem. Res. Toxicol.

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“…Because of the site-selectivity inherent in the method, it has been successfully used to identify loci of damage in DNA produced by addition reactions with carcinogens derived from polycyclic aromatic hydrocarbons [8].…”

Section: Improvement Of Selectivity Based Upon Narrow-band Excitationmentioning

confidence: 99%

Fluorescence Spectroscopy: Where We Are and Where We’re Going

Schulman

1993

Fluorescence Spectroscopy

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“…Particularly powerful is high-resolution, fluorescence line-narrowing spectroscopy (FLNS), attributed with fingerprinting capabilities and proven to provide unambiguous identification of structurally related fluorescent analytes, i.e., isomers (8,9). Its physical principles and attributes are only briefly mentioned here; for a detailed description, the reader is referred to recent reviews and book chapters (10,11).…”

mentioning

confidence: 99%

Cross-reactivity and conformational multiplicity of an anti-polycyclic aromatic hydrocarbon mAb

Grubor

Hayes²,

Small³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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Cross-reactivity and multispecific functionality of antibodies play a central role in the immune system. The Ab's promiscuity is attributed to structural flexibility and conformational multiplicity of its binding sites governed by the rearrangement of hydrogen bonding centers. However, antibodies whose recognition and binding rely on less directional hydrophobic interactions might follow different interaction pathways. We investigated interaction of anti-polycyclic aromatic hydrocarbon mAb with two biologically important cross-reactants, pyrene and benzo(a)pyrene. Complex formation was characterized by means of low-temperature laser-induced fluorescence spectroscopy in both low-and high-resolution fluorescence line-narrowing (FLN) modes. It is shown that the FLN spectroscopy can identify various haptens cross-reacted with an Ab, as well as simultaneously differentiate between free and immunocomplexed haptens. In addition, our results suggest an interesting case of an Ab binding a particular cross-reactant by adopting two distinct conformations of its binding sites. The existence of the multiple conformations for anti-polycyclic aromatic hydrocarbon mAb that are trapped at low temperature can be rationalized through the existing models for Ab binding. Finally, as revealed by FLN spectra of immunocomplexed chromophores, -interactions, rather than hydrogen bonding, play the central role in complex formation.fluorescence spectroscopy T he relationship between protein structure and function continues to be one of the most challenging of problems in molecular biology. Complicating the solution of this problem is the demise of the view that a given protein sequence has one structure and, therefore, one function. Indeed, examples of multifunctional proteins are numerous, as are examples in which a single binding site can bind a range of varied molecular shapes (1-3). This promiscuity of proteins has been explained by two different models: the molecular mimicry model and preexisting equilibrium of structural isomers. The molecular mimicry model assumes that the protein exists in a single conformation that can bind structurally similar ligands with low affinity and that, subsequently, the protein rearranges into a high-affinity configuration (induced fit). Evidence for the induced fit model is that crystal structures of bound and unbound proteins show differences in the ligand-binding site configuration (2, 4). The preexisting equilibrium model, on the other hand, assumes that the intact protein exists in multiple conformations, one of which has high affinity for the ligand; following binding, the equilibrium shifts in favor of this configuration (5). Recently, James and colleagues (2, 3) presented structural evidence for equilibrium between the two preexisting Ab isomers and demonstrated that a single Ab (SPE7, raised against 2,4-dinitrophenyl hapten) can bind structurally unrelated ligands through conformational selection (3). They have also investigated cross-reactivity of the same Ab involving a single conformational ...

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Fluorescence-Line-Narrowed Spectra of Polycyclic Aromatic Carcinogen-DNA Adducts

Cited by 29 publications

References 10 publications

Fluorescence line narrowing-nonphotochemical hole burning spectrometry: femtomole detection and high selectivity for intact DNA-PAH adducts

Fluorescence line narrowing-nonphotochemical hole burning spectrometry: femtomole detection and high selectivity for intact DNA-PAH adducts

Fluorescence Spectroscopy: Where We Are and Where We’re Going

Cross-reactivity and conformational multiplicity of an anti-polycyclic aromatic hydrocarbon mAb

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