Some experiments with spectrofluorimeters and filter fluorimeters

Parker, C. A.; Barnes, W. Jon. P.

doi:10.1039/an9578200606

Cited by 214 publications

(100 citation statements)

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“…KI quenching data were corrected for ionic strength effects by parallel titrations with potassium chloride; to correct for the inner filter effect of acrylamide, the formula of Parker and Barnes (1957) was applied. The data were plotted according to the Stern-Volmer equation,…”

Section: Fluorescence Spectroscopymentioning

confidence: 99%

The tryptophan residues of mitochondrial creatine kinase: Roles of Trp‐223, Trp‐206, and Trp‐264 in active‐site and quaternary structure formation

et al. 1994

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The 5 tryptophan residues of chicken sarcomeric mitochondrial creatine kinase (Mi,-CK) were individually replaced by phenylalanine or cysteine using site-directed mutagenesis. The mutant proteins were analyzed by enzyme kinetics, fluorescence spectroscopy, circular dichroism, and conformational stability studies. In the present work, Trp-223 is identified as an active-site residue whose replacement even by phenylalanine resulted in 296% inactivation of the enzyme. Trp-223 is responsible for a strong (18-21%) fluorescence quenching effect occurring upon formation of a transition state-analogue complex (TSAC; Mib-CK .creatine.MgADP .NO,-), and Trp-223 is probably required for the conformational change leading to the TSAC-induced octamer dissociation of Mib-CK.Replacement of Trp-206 by cysteine led to a destabilization of the active-site structure, solvent exposure of Trp-223, and to the dissociation of the Mib-CK dimers into monomers. However, this dimer dissociation was counteracted by TSAC formation or the presence of ADP alone. Trp-264 is shown to be located at the dimerdimer interfaces within the Mib-CK octamer, being the origin of another strong (25%) fluorescence quenching effect, which was observed upon the TSAC-induced octamer dissociation. Substitution of Trp-264 by cysteine drastically accelerated the TSAC-induced dissociation and destabilized the octameric structure by one-fourth of the total free interaction energy, probably by weakening hydrophobic contacts. The roles of the other 2 tryptophan residues, Trp-213 and Trp-268, could be less well assigned.

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Section: Fluorescence Spectroscopymentioning

confidence: 99%

The tryptophan residues of mitochondrial creatine kinase: Roles of Trp‐223, Trp‐206, and Trp‐264 in active‐site and quaternary structure formation

et al. 1994

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“…Attenuation of the excitation beam before reaching the region viewed by the fluorescence detection optics (pre-filter region) and through the interrogation volume element is denoted as primary inner-filtering. The correction factor, fprimt for primary inner-filtering is given by the following expression: 45 After the initial intensity reading, unless otherwise specified, each successive value corresponds to addition of one pasteur pipet drop of nitromethane to the PANH dissolved in aqueous acetonitrile Table I for the 14 alternant PANH solutes studied. Careful examination reveals that pobs foes decrease with increasing nitromethane concen tration, though not all of the intensity reduction is at tributed to fluorescence quenching.…”

Section: Discussion Of Fluorescence Quenching Resultsmentioning

confidence: 99%

Spectroscopic Properties of Polycyclic Aromatic Compounds: Examination of Nitromethane as a Selective Fluorescence Quenching Agent for Alternant Polycyclic Aromatic Nitrogen Hetero-Atom Derivatives

et al. 1992

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Nitromethane is examined as a selective fluorescence quenching agent for alternant polycyclic aromatic nitrogen hetero-atoms (PANHs). Fluorescence emission behavior is reported for 1-azapyrene, 2-azapyrene, 4-azapyrene, 4-azachrysene, 12-azabenzo[a]pyrene, phenanthro[9, 10g]isoquinoline, phenanthro[2,3h]isoquinoline, phenanthro[3,2h]-isoquinoline, 2-azabenz[a]anthracene, 1-azabenz[a]anthracene, 9-azabenz[a]anthracene, dibenzo[c,i]phenanthro[1,10,9,8anmlk]phenanthridine, diphenanthro[9,10,1def;1',10',9'hij]phthalazine, and benz[de]isoquino[1,8gh]quinoline dissolved in acetonitrile or aqueous-acetonitrile solvent mixtures at various nitromethane concentrations. Results of these measurements show that nitromethane does quench fluorescence emission of ten of the solutes studied; however, phenanthro-[2,3h]isoquinoline, 9-azabenz[a]anthracene, benz[de]isoquino[1,8gh]-quinoline, and dibenzo[c,i]phenanthro[1,10,9,8anmlk]phenanthridine are notable exceptions.

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“…No correction was needed, therefore, for the inner filter absorption by the solution. Both phosphorescence excitation and fluorescence excitation spectra were corrected for the spectral output of the Xe lamp and the corresponding spectral dispersion a t various wavelengths for the slit widths employed (11)(12)(13)(14).…”

Section: Organic Emission Of Free Ligands and Gd Chelates And Phosphomentioning

confidence: 99%

Energy transfer in mixed lanthanide complexes

Filipescu¹,

Bjorklund²,

Mcavoy³

et al. 1967

Can. J. Chem.

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Some experiments with spectrofluorimeters and filter fluorimeters

Cited by 214 publications

References 0 publications

The tryptophan residues of mitochondrial creatine kinase: Roles of Trp‐223, Trp‐206, and Trp‐264 in active‐site and quaternary structure formation

The tryptophan residues of mitochondrial creatine kinase: Roles of Trp‐223, Trp‐206, and Trp‐264 in active‐site and quaternary structure formation

Spectroscopic Properties of Polycyclic Aromatic Compounds: Examination of Nitromethane as a Selective Fluorescence Quenching Agent for Alternant Polycyclic Aromatic Nitrogen Hetero-Atom Derivatives

Energy transfer in mixed lanthanide complexes

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