In silico characterization of resonance energy transfer for disk-shaped membrane domains
In silico characterization of resonance energy transfer for disk-shaped membrane domains
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DOI：
10.1529/biophysj.106.093245
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发表时间：
2007-05-01
影响因子：
3.4
通讯作者：
Kenworthy, Anne K.
中科院分区：
文献类型：
--
作者：
Kiskowski, Maria A.;Kenworthy, Anne K.
福斯特共振能量转移（FRET）已成为研究膜中蛋白质和脂质亚微米分布的重要工具。虽然从 FRET 中解析荧光团的二维分布通常是不确定的，但可以使用正向方法来确定感兴趣的微域组织类别的特征 FRET“签名”。作为实现这一目标的第一步，我们使用随机蒙特卡罗方法来表征分子随机分布在盘形域内的情况下的 FRET。我们发现，当供体和受体被限制在域内时，FRET 通常取决于域内受体的密度。结果是，即使结构域具有不同的直径或分子数量，也无法通过这种 FRET 方法区分结构域直径和分子数量。相反，可以通过将此方法与测量限制在结构域内的供体和位于结构域外的受体之间的 FRET 的分离方法相结合来解决。这些发现界定了此类分布的逆问题的可处理性，并重新构建了 FRET 可用于表征微结构域（例如脂筏）结构的方式。
Forster resonance energy transfer (FRET) has become an important tool to study the submicrometer distribution of proteins and lipids in membranes. Although resolving the two-dimensional distribution of fluorophores from FRET is generally underdetermined, a forward approach can be used to determine characteristic FRET "signatures'' for interesting classes of microdomain organizations. As a first step toward this goal, we use a stochastic Monte Carlo approach to characterize FRET in the case of molecules randomly distributed within disk-shaped domains. We find that when donors and acceptors are confined within domains, FRET depends very generally on the density of acceptors within domains. An implication of this result is that two domain populations with the same acceptor density cannot be distinguished by this FRET approach even if the domains have different diameters or different numbers of molecules. In contrast, both the domain diameter and molecule number can be resolved by combining this approach with a segregation approach that measures FRET between donors confined in domains and acceptors localized outside domains. These findings delimit where the inverse problem is tractable for this class of distributions and reframe ways FRET can be used to characterize the structure of microdomains such as lipid rafts.