Quantitative single-molecule conformational distributions

A case study with poly-(L-proline)

Lucas P. Watkins, Hauyee Chang, Haw Yang

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

Precise measurement of the potential of mean force is necessary for a fundamental understanding of the dynamics and chemical reactivity of a biological macromolecule. The unique advantage provided by the recently developed constant-information approach to analyzing time-dependent single-molecule fluorescence measurements was used with maximum entropy deconvolution to create a procedure for the accurate determination of molecular conformational distributions, and analytical expressions for the errors in these distributions were derived. This new method was applied to a derivatized poly(L-proline) series, P nCG 3K-(biotin) (n = 8, 12, 15, 18, and 24), using a modular, server-based single-molecule spectrometer that is capable of registering photon arrival times with a continuous-wave excitation source. To account for potential influence from the microscopic environment, factors that were calibrated and corrected molecule by molecule include background, cross-talk, and detection efficiency. For each single poly(L-proline) molecule, sharply peaked Förster type resonance energy transfer (FRET) efficiency and distance distributions were recovered, indicating a static end-to-end distance on the time scale of measurement. The experimental distances were compared with models of varying rigidity. The results suggest that the 23 A persistence length wormlike chain model derived from experiments with high molecular weight poly(L-proline) is applicable to short chains as well.

Original languageEnglish (US)
Pages (from-to)5191-5203
Number of pages13
JournalJournal of Physical Chemistry A
Volume110
Issue number15
DOIs
StatePublished - Apr 20 2006
Externally publishedYes

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Molecules
molecules
Chemical reactivity
biotin
wave excitation
Deconvolution
Biotin
Macromolecules
Chain length
macromolecules
rigidity
Rigidity
Energy transfer
continuous radiation
arrivals
Spectrometers
molecular weight
Entropy
Servers
Photons

All Science Journal Classification (ASJC) codes

  • Physical and Theoretical Chemistry

Cite this

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Quantitative single-molecule conformational distributions : A case study with poly-(L-proline). / Watkins, Lucas P.; Chang, Hauyee; Yang, Haw.

In: Journal of Physical Chemistry A, Vol. 110, No. 15, 20.04.2006, p. 5191-5203.

Research output: Contribution to journalArticle

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