Refining membrane penetration by a combination of steady-state and time-resolved depth-dependent fluorescence quenching

被引：12

作者：

Kyrychenko, Alexander ^{[1
]}

Ladokhin, Alexey S. ^{[1
]}

机构：

[1] Univ Kansas, Med Ctr, Dept Biochem & Mol Biol, Kansas City, KS 66160 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2014年 / 446卷

基金：

美国国家卫生研究院;

关键词：

Static and dynamic fluorescence quenching; Bilayer penetration; Distribution analysis; Lipid-attached quenchers; SPIN-LABELED PHOSPHOLIPIDS; TRYPTOPHAN OCTYL ESTER; MODEL MEMBRANES; PROTEINS; DYNAMICS; BILAYER; ORIENTATION; PEPTIDES; TOPOLOGY; RESIDUES;

D O I：

10.1016/j.ab.2013.10.015

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Accurate determination of the depth of membrane penetration of a fluorescent probe, attached to a lipid, protein, or other macromolecule of interest, using depth-dependent quenching methodology is complicated by thermal motion in the lipid bilayer. Here, we suggest that a combination of steady-state and time-resolved measurements can be used to generate a static quenching profile that reduces the contribution from transverse diffusion occurring during the excited-state lifetime. This procedure results in narrower quenching profiles, compared with those obtained by traditional measurements, and thus improves precision in determination of the underlying depth distribution of the probe. (C) 2013 Elsevier Inc. All rights reserved.

引用

页码：19 / 21

页数：3

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