Determination of intramolecular distance distribution during protein folding on the millisecond timescale

Vladimir Ratner; Michael Sinev; Elisha Haas

doi:10.1006/jmbi.2000.3814

Determination of intramolecular distance distribution during protein folding on the millisecond timescale

Vladimir Ratner, Michael Sinev, Elisha Haas

The Mina and Everard Goodman Faculty of Life Sciences

Bar-Ilan University

Research output: Contribution to journal › Article › peer-review

46 Scopus citations

Abstract

A method for determination of transient (on the millisecond timescale) intramolecular distance distributions (IDDs) by time-resolved dynamic non-radiative excitation energy transfer measurements was developed. The time-course of the development of the IDD between residues 73 and 203 in the CORE domain of Escherichia coli adenylate kinase throughout refolding from the GuHCl-induced denatured state was determined. The mean of the apparent IDD reduced to a value close to its magnitude in the native protein, within 2 ms (the dead-time of the instrument). At that time the width of that distribution was rather large (16 ± 2 Å). The large width implies that the intramolecular diffusion coefficient of the labeled segment does not exceed 10^-7 cm²/second. In a second slower phase of the refolding transition, the width was reduced to its native value (6 ± 4 Å). (C) 2000 Academic Press.

Original language	English
Pages (from-to)	1363-1371
Number of pages	9
Journal	Journal of Molecular Biology
Volume	299
Issue number	5
DOIs	https://doi.org/10.1006/jmbi.2000.3814
State	Published - 23 Jun 2000

Bibliographical note

Funding Information:
We thank E. Sineva for preparation of mutant plasmids, J. Shwartzburd for protein purification, Dr E. Cahana for technical assistance, A. Goldin for supplying software, Professor E. Meirovicth and Professor W. B. Gratzer of Kings College London for helpful comments on the manuscript. This research was supported by grants from the Israel Science Foundation, the Israel Ministry of Science and the National Institute of Health (USA) RO1-GM39372.

Funding

We thank E. Sineva for preparation of mutant plasmids, J. Shwartzburd for protein purification, Dr E. Cahana for technical assistance, A. Goldin for supplying software, Professor E. Meirovicth and Professor W. B. Gratzer of Kings College London for helpful comments on the manuscript. This research was supported by grants from the Israel Science Foundation, the Israel Ministry of Science and the National Institute of Health (USA) RO1-GM39372.

Funders	Funder number
National institute of Health	RO1-GM39372
National Institute of General Medical Sciences	R01GM039372
Israel Science Foundation
Ministry of science and technology, Israel

Keywords

Adenylate kinase
Distance distribution
Energy transfer
Folding
Time-resolved

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10.1006/jmbi.2000.3814

Cite this

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abstract = "A method for determination of transient (on the millisecond timescale) intramolecular distance distributions (IDDs) by time-resolved dynamic non-radiative excitation energy transfer measurements was developed. The time-course of the development of the IDD between residues 73 and 203 in the CORE domain of Escherichia coli adenylate kinase throughout refolding from the GuHCl-induced denatured state was determined. The mean of the apparent IDD reduced to a value close to its magnitude in the native protein, within 2 ms (the dead-time of the instrument). At that time the width of that distribution was rather large (16 ± 2 {\AA}). The large width implies that the intramolecular diffusion coefficient of the labeled segment does not exceed 10-7 cm2/second. In a second slower phase of the refolding transition, the width was reduced to its native value (6 ± 4 {\AA}). (C) 2000 Academic Press.",

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N2 - A method for determination of transient (on the millisecond timescale) intramolecular distance distributions (IDDs) by time-resolved dynamic non-radiative excitation energy transfer measurements was developed. The time-course of the development of the IDD between residues 73 and 203 in the CORE domain of Escherichia coli adenylate kinase throughout refolding from the GuHCl-induced denatured state was determined. The mean of the apparent IDD reduced to a value close to its magnitude in the native protein, within 2 ms (the dead-time of the instrument). At that time the width of that distribution was rather large (16 ± 2 Å). The large width implies that the intramolecular diffusion coefficient of the labeled segment does not exceed 10-7 cm2/second. In a second slower phase of the refolding transition, the width was reduced to its native value (6 ± 4 Å). (C) 2000 Academic Press.

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Determination of intramolecular distance distribution during protein folding on the millisecond timescale

Abstract

Bibliographical note

Funding

Keywords

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