Measurement of subnanosecond anisotropy decays of protein fluorescence using frequency-domain fluorometry

J. R. Lakowicz, G. Laczko, Ignacy Gryczynski, H. Cherek

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

We report the first anisotropy decays of protein fluorescence obtained using a frequency-domain fluorometer. The ultraviolet light source (300 nm) was a ring dye laser equipped with an intracavity frequency doubler, pumped by an argon ion laser. The data, measured at modulation frequencies from 2 to 200 MHz, reveal the presence of subnanosecond motions (0.1-0.2 ns) of the single tryptophan residues in melittin and monellin. For melittin the data also indicate the presence of slower motions near 1 ns, which may be the result of concerted motions of several peptide units. Smaller amplitude motions, on a similar timescale, were observed for the single tryptophan residue in staphylococcal nuclease. We demonstrate using N-acetyl-L-tryptophanamide in water that the method of frequency-domain fluorometry is capable of measuring correlation times as short as 50 ps. This method can provide data for the direct comparison of measured anisotropy decays with those prediced from molecular dynamics calculations.

Original languageEnglish
Pages (from-to)2240-2245
Number of pages6
JournalJournal of Biological Chemistry
Volume261
Issue number5
StatePublished - 1 Dec 1986

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Melitten
Fluorometry
Anisotropy
Tryptophan
Fluorescence
Fluorometers
Frequency doublers
Micrococcal Nuclease
Ring lasers
Dye lasers
Argon
Frequency modulation
Light sources
Molecular dynamics
Proteins
Ions
Dye Lasers
Peptides
Gas Lasers
Water

Cite this

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abstract = "We report the first anisotropy decays of protein fluorescence obtained using a frequency-domain fluorometer. The ultraviolet light source (300 nm) was a ring dye laser equipped with an intracavity frequency doubler, pumped by an argon ion laser. The data, measured at modulation frequencies from 2 to 200 MHz, reveal the presence of subnanosecond motions (0.1-0.2 ns) of the single tryptophan residues in melittin and monellin. For melittin the data also indicate the presence of slower motions near 1 ns, which may be the result of concerted motions of several peptide units. Smaller amplitude motions, on a similar timescale, were observed for the single tryptophan residue in staphylococcal nuclease. We demonstrate using N-acetyl-L-tryptophanamide in water that the method of frequency-domain fluorometry is capable of measuring correlation times as short as 50 ps. This method can provide data for the direct comparison of measured anisotropy decays with those prediced from molecular dynamics calculations.",
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Measurement of subnanosecond anisotropy decays of protein fluorescence using frequency-domain fluorometry. / Lakowicz, J. R.; Laczko, G.; Gryczynski, Ignacy; Cherek, H.

In: Journal of Biological Chemistry, Vol. 261, No. 5, 01.12.1986, p. 2240-2245.

Research output: Contribution to journalArticle

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AU - Lakowicz, J. R.

AU - Laczko, G.

AU - Gryczynski, Ignacy

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N2 - We report the first anisotropy decays of protein fluorescence obtained using a frequency-domain fluorometer. The ultraviolet light source (300 nm) was a ring dye laser equipped with an intracavity frequency doubler, pumped by an argon ion laser. The data, measured at modulation frequencies from 2 to 200 MHz, reveal the presence of subnanosecond motions (0.1-0.2 ns) of the single tryptophan residues in melittin and monellin. For melittin the data also indicate the presence of slower motions near 1 ns, which may be the result of concerted motions of several peptide units. Smaller amplitude motions, on a similar timescale, were observed for the single tryptophan residue in staphylococcal nuclease. We demonstrate using N-acetyl-L-tryptophanamide in water that the method of frequency-domain fluorometry is capable of measuring correlation times as short as 50 ps. This method can provide data for the direct comparison of measured anisotropy decays with those prediced from molecular dynamics calculations.

AB - We report the first anisotropy decays of protein fluorescence obtained using a frequency-domain fluorometer. The ultraviolet light source (300 nm) was a ring dye laser equipped with an intracavity frequency doubler, pumped by an argon ion laser. The data, measured at modulation frequencies from 2 to 200 MHz, reveal the presence of subnanosecond motions (0.1-0.2 ns) of the single tryptophan residues in melittin and monellin. For melittin the data also indicate the presence of slower motions near 1 ns, which may be the result of concerted motions of several peptide units. Smaller amplitude motions, on a similar timescale, were observed for the single tryptophan residue in staphylococcal nuclease. We demonstrate using N-acetyl-L-tryptophanamide in water that the method of frequency-domain fluorometry is capable of measuring correlation times as short as 50 ps. This method can provide data for the direct comparison of measured anisotropy decays with those prediced from molecular dynamics calculations.

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