ANISOTROPY DECAYS of SINGLE TRYPTOPHAN PROTEINS MEASURED by GHz FREQUENCY-DOMAIN FLUOROMETRY with COLLISIONAL QUENCHING

Joseph R. Lakowicz; Ignacy Gryczynski; Henryk Szmacinski; Henryk Cherek; Nanda Joshi

doi:10.1117/12.945384

ANISOTROPY DECAYS of SINGLE TRYPTOPHAN PROTEINS MEASURED by GHz FREQUENCY-DOMAIN FLUOROMETRY with COLLISIONAL QUENCHING

Joseph R. Lakowicz, Ignacy Gryczynski, Henryk Szmacinski, Henryk Cherek, Nanda Joshi

Research output: Contribution to journal › Conference article › peer-review

1 Scopus citations

Abstract

We use frequency-domain fluorometry to determine the anisotropy decays of the tryptophan emission from S. nuclease and from the model compound gly-trp-gly. Resolution of the rapid and complex anisotropy decays was enhanced by global analysis of the data measured in the presence of quenching by either oxygen or acrylamide. Data were obtained at four to six quencher concentrations, and the data analyzed globally to recover the anisotropy decay. Because the decay times were decreased by quenching, measurements were possible to a upper frequency limit of 2 GHz. The anisotropy decay of gly-trp-gly revealed 40 ps of the indole ring, which was resolved from the overall 150 ps correlation times of the tripeptide. The anisotropy decay of nuclease displayed a 90 /ps component as well as a 10 ns component due to overall rotational diffusion. We believe these highly resolved anisotropy decays are suitably for comparison with molecular dynamic simulations.

Original language	English
Pages (from-to)	178-185
Number of pages	8
Journal	Proceedings of SPIE - The International Society for Optical Engineering
Volume	909
DOIs	https://doi.org/10.1117/12.945384
State	Published - 24 Jun 1988
Event	Time-Resolved Laser Spectroscopy in Biochemistry 1988 - Los Angeles, United States Duration: 11 Jan 1988 → 17 Jan 1988

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@article{7544c17706f248f9940e657f92434573,

title = "ANISOTROPY DECAYS of SINGLE TRYPTOPHAN PROTEINS MEASURED by GHz FREQUENCY-DOMAIN FLUOROMETRY with COLLISIONAL QUENCHING",

abstract = "We use frequency-domain fluorometry to determine the anisotropy decays of the tryptophan emission from S. nuclease and from the model compound gly-trp-gly. Resolution of the rapid and complex anisotropy decays was enhanced by global analysis of the data measured in the presence of quenching by either oxygen or acrylamide. Data were obtained at four to six quencher concentrations, and the data analyzed globally to recover the anisotropy decay. Because the decay times were decreased by quenching, measurements were possible to a upper frequency limit of 2 GHz. The anisotropy decay of gly-trp-gly revealed 40 ps of the indole ring, which was resolved from the overall 150 ps correlation times of the tripeptide. The anisotropy decay of nuclease displayed a 90 /ps component as well as a 10 ns component due to overall rotational diffusion. We believe these highly resolved anisotropy decays are suitably for comparison with molecular dynamic simulations.",

author = "Lakowicz, {Joseph R.} and Ignacy Gryczynski and Henryk Szmacinski and Henryk Cherek and Nanda Joshi",

year = "1988",

month = jun,

day = "24",

doi = "10.1117/12.945384",

language = "English",

volume = "909",

pages = "178--185",

journal = "Proceedings of SPIE - The International Society for Optical Engineering",

issn = "0277-786X",

publisher = "SPIE",

note = "null ; Conference date: 11-01-1988 Through 17-01-1988",

}

ANISOTROPY DECAYS of SINGLE TRYPTOPHAN PROTEINS MEASURED by GHz FREQUENCY-DOMAIN FLUOROMETRY with COLLISIONAL QUENCHING. / Lakowicz, Joseph R.; Gryczynski, Ignacy; Szmacinski, Henryk; Cherek, Henryk; Joshi, Nanda.

In: Proceedings of SPIE - The International Society for Optical Engineering, Vol. 909, 24.06.1988, p. 178-185.

Research output: Contribution to journal › Conference article › peer-review

TY - JOUR

T1 - ANISOTROPY DECAYS of SINGLE TRYPTOPHAN PROTEINS MEASURED by GHz FREQUENCY-DOMAIN FLUOROMETRY with COLLISIONAL QUENCHING

AU - Lakowicz, Joseph R.

AU - Gryczynski, Ignacy

AU - Szmacinski, Henryk

AU - Cherek, Henryk

AU - Joshi, Nanda

PY - 1988/6/24

Y1 - 1988/6/24

N2 - We use frequency-domain fluorometry to determine the anisotropy decays of the tryptophan emission from S. nuclease and from the model compound gly-trp-gly. Resolution of the rapid and complex anisotropy decays was enhanced by global analysis of the data measured in the presence of quenching by either oxygen or acrylamide. Data were obtained at four to six quencher concentrations, and the data analyzed globally to recover the anisotropy decay. Because the decay times were decreased by quenching, measurements were possible to a upper frequency limit of 2 GHz. The anisotropy decay of gly-trp-gly revealed 40 ps of the indole ring, which was resolved from the overall 150 ps correlation times of the tripeptide. The anisotropy decay of nuclease displayed a 90 /ps component as well as a 10 ns component due to overall rotational diffusion. We believe these highly resolved anisotropy decays are suitably for comparison with molecular dynamic simulations.

AB - We use frequency-domain fluorometry to determine the anisotropy decays of the tryptophan emission from S. nuclease and from the model compound gly-trp-gly. Resolution of the rapid and complex anisotropy decays was enhanced by global analysis of the data measured in the presence of quenching by either oxygen or acrylamide. Data were obtained at four to six quencher concentrations, and the data analyzed globally to recover the anisotropy decay. Because the decay times were decreased by quenching, measurements were possible to a upper frequency limit of 2 GHz. The anisotropy decay of gly-trp-gly revealed 40 ps of the indole ring, which was resolved from the overall 150 ps correlation times of the tripeptide. The anisotropy decay of nuclease displayed a 90 /ps component as well as a 10 ns component due to overall rotational diffusion. We believe these highly resolved anisotropy decays are suitably for comparison with molecular dynamic simulations.

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U2 - 10.1117/12.945384

DO - 10.1117/12.945384

M3 - Conference article

AN - SCOPUS:84913484403

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EP - 185

JO - Proceedings of SPIE - The International Society for Optical Engineering

JF - Proceedings of SPIE - The International Society for Optical Engineering

SN - 0277-786X

Y2 - 11 January 1988 through 17 January 1988

ER -

ANISOTROPY DECAYS of SINGLE TRYPTOPHAN PROTEINS MEASURED by GHz FREQUENCY-DOMAIN FLUOROMETRY with COLLISIONAL QUENCHING

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