Kinetics of fast changing intramolecular distance distributions obtained by combined analysis of FRET efficiency kinetics and time-resolved FRET equilibrium measurements

E. Lerner, T. Orevi, E. Ben Ishay, D. Amir, E. Haas

نتاج البحث: نشر في مجلةمقالةمراجعة النظراء

ملخص

Detailed studies of the mechanisms of macromolecular conformational transitions such as protein folding are enhanced by analysis of changes of distributions for intramolecular distances during the transitions. Time-resolved Förster resonance energy transfer (FRET) measurements yield such data, but the more readily available kinetics of mean FRET efficiency changes cannot be analyzed in terms of changes in distances because of the sixth-power dependence on the mean distance. To enhance the information obtained from mean FRET efficiency kinetics, we combined the analyses of FRET efficiency kinetics and equilibrium trFRET experiments. The joint analysis enabled determination of transient distance distributions along the folding reaction both in cases where a two-state transition is valid and in some cases consisting of a three-state scenario. The procedure and its limits were tested by simulations. Experimental data obtained from stopped-flow measurements of the refolding of Escherichia coli adenylate kinase were analyzed. The distance distributions between three double-labeled mutants, in the collapsed transient state, were determined and compared to those obtained experimentally using the double-kinetics technique. The proposed method effectively provides information on distance distributions of kinetically accessed intermediates of fast conformational transitions induced by common relaxation methods.

اللغة الأصليةإنجليزيّة أمريكيّة
الصفحات (من إلى)667-676
عدد الصفحات10
دوريةBiophysical Journal
مستوى الصوت106
رقم الإصدار3
المعرِّفات الرقمية للأشياء
حالة النشرنُشِر - 4 فبراير 2014

All Science Journal Classification (ASJC) codes

  • !!Biophysics

بصمة

أدرس بدقة موضوعات البحث “Kinetics of fast changing intramolecular distance distributions obtained by combined analysis of FRET efficiency kinetics and time-resolved FRET equilibrium measurements'. فهما يشكلان معًا بصمة فريدة.

قم بذكر هذا