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

Vladimir Ratner, Michael Sinev, Elisha Haas

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46 Scopus citations


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 languageEnglish
Pages (from-to)1363-1371
Number of pages9
JournalJournal of Molecular Biology
Issue number5
StatePublished - 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.


  • Adenylate kinase
  • Distance distribution
  • Energy transfer
  • Folding
  • Time-resolved


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