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J Phys Chem B. 2018 Apr 26;122(16):4445-4456. doi: 10.1021/acs.jpcb.8b00879. Epub 2018 Apr 17.

Method-Unifying View of Loop-Formation Kinetics in Peptide and Protein Folding.

Author information

1
Department of Life Sciences and Chemistry , Jacobs University Bremen , Bremen 28759 , Germany.
2
Center for Biotechnology and Bioengineering , China University of Petroleum , Qingdao , Shandong , China 266580.
3
Department of Life Science , Bar Ilan University , Ramat Gan 5290002 , Israel.

Abstract

Protein folding can be described as a probabilistic succession of events in which the peptide chain forms loops closed by specific amino acid residue contacts, herein referred to as loop nodes. To measure loop rates, several photophysical methods have been introduced where a pair of optically active probes is incorporated at selected chain positions and the excited probe undergoes contact quenching (CQ) upon collision with the second probe. The quenching mechanisms involved triplet-triplet energy transfer, photoinduced electron transfer, and collision-induced fluorescence quenching, where the fluorescence of Dbo, an asparagine residue conjugated to 2,3-diazabicyclo[2.2.2]octane, is quenched by tryptophan. The discrepancy between the loop rates afforded from these three CQ techniques has, however, remained unresolved. In analyzing this discrepancy, we now report two short-distance FRET methods where Dbo acts as an energy acceptor in combination with tryptophan and naphtylalanine, two donors with largely different fluorescence lifetimes of 1.3 and 33 ns, respectively. Despite the different quenching mechanisms, the rates from FRET and CQ methods were, surprisingly, of comparable magnitude. This combination of FRET and CQ data led to a unifying physical model and to the conclusion that the rate of loop formation in folding reactions varies not only with the kind and number of residues that constitute the chain but also in particular with the size and properties of the residues that constitute the loop node.

PMID:
29617564
DOI:
10.1021/acs.jpcb.8b00879
[Indexed for MEDLINE]

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