Hindered depolarizing rotations of perylene in lipid bilayers. Detection by lifetime-resolved fluorescence anisotropy measurements.
Hindered depolarizing rotations of perylene in lipid bilayers. Detection by lifetime-resolved fluorescence anisotropy measurements.
复制标题
阻碍脂质双层中苝的去极化旋转。
DOI：
10.1021/bi00546a013
复制
发表时间：
1980
期刊：
影响因子：
2.9
通讯作者：
Knutson,JR
中科院分区：
文献类型：
--
作者：
Lakowicz,JR;Knutson,JR
J·R·Lakowicz和Jay R·Knutson摘要：利用分子氧猝灭分子的荧光来改变其荧光寿命。在这些猝灭条件下的稳态荧光各向异性测量被用来研究二茂铁在各向同性溶剂丙二醇中和在脂质双层中的扩散运动。这些寿命分辨的各向异性测量表明，与其荧光寿命相比，丙二醇组分的各向异性在更长的时间内衰减到零。相反，在DMPC囊泡中，在这些长时间(r“)处的渐近或”极限“各向异性值不为零。/·”值在低于DMPC相变温度23C时最大。在5℃和47℃时，DMPC囊泡中Perene的代表值分别为0.16和0.02。因此，与在IVI中观察到的沸石的自由旋转相反，膜结合荧光团的稳态荧光各向异性的测量已被广泛用于估计脂双层(Shinitzky等人，1971；Cogan等人，1973；Lentz等人，1976；Brashford等人，1976；Jacobson&Wob-Schall，1974)和生物膜(Moore等人，1976；Helgerson等人，1974)的酰基侧链区域的“微粘度”。在从观察到的荧光各向异性到膜的微粘性值的外推中，人们假设在膜中和在参考溶剂中荧光团的去极化旋转的性质是相同的。我们所说的自然是指这些旋转的自由和各向同性。最近，对于广泛使用的微粘度探针二苯基己三烯(DPH)1**(Kawato等人，1977；Veatch&Stryer，1977；Dale等人，1977；Lakowicz&Prendergast，1978；Lakowicz等人，1979a，b)、2-苯胺基-8-萘磺酸(Badea等人，1978)和1-苯胺基-8-萘磺酸(Kinosita等人，1976)，这种假设被证明是错误的。对于这些探针中的每一个，发现膜中的扩散运动受到阻碍，与荧光寿命(r“)相比，在时间上存在非零的各向异性值就是证据。我们现在提出的证据表明，另一种广泛使用的微粘度探针，聚乙烯，当结合到脂质双层时，也是一个受阻的旋转体。事实上，决定传统测量的稳态各向异性的是它的自转受阻程度，而不是它的自转速度。这些观察结果表明，在从各向异性测量得出的膜微粘度的字面解释中需要谨慎。
J. R. Lakowicz* and Jay R. Knutson abstract: Oxygen quenching of perylene fluorescence was used to vary its fluorescence lifetime. Steady-state fluorescence anisotropy measurements under these quenchingconditions were used to investigate the diffusive motions of perylene in the isotropic solvent propylene glycol and in lipid bilayers. These lifetime-resolved anisotropy measurements indicate that the anisotropy of perylene in propylene glycol decays to zero at times long compared to its fluorescence lifetime. In contrast, the asymptotic or “limiting” anisotropy values atthese long times (r „) are nonzero in vesicles of dimyristoyl-phosphatidylcholine (DMPC)./·„values are largest at tem-peratures below the DMPC phase transition temperature of 23 C. Representative values of for perylene in DMPC vesicles are 0.16 and 0.02 at 5 and 47 C, respectively. Thus, in contrast to the free rotations observed for perylene inIVIeasurements of the steady-state fluorescence anisotropies of membrane-bound fluorophores have become widely used in estimating the “microviscosity” of the acyl side chain region of lipid bilayers (Shinitzky et al., 1971; Cogan et al., 1973; Lentz et al., 1976; Brashford et al., 1976; Jacobson & Wob-schall, 1974) and of biological membranes (Moore et al., 1976; Helgerson et al., 1974). In the extrapolation from an observed fluorescence anisotropy to a microviscosity value for the membrane, one assumes that the nature of the depolarizing rotations of the fluorophore in the membrane and in the reference solvent is identical. By nature we mean the freedom and isotropy of these rotations. Recently this assumption was shown to be in error for the widely used microviscosity probe diphenylhexatriene (DPH) 1**(Kawato etal., 1977; Veatch & Stryer, 1977; Dale et al., 1977; Lakowicz & Prendergast, 1978; Lakowicz et al., 1979a, b), for 2-anilinonaphthalene (Badea et al., 1978), and for 1-anilino-8-naphthalenesulfonic acid (Kinosita et al., 1976). For each of these probes the diffusive motions in membranes were found to be hindered, as evidenced by the existence of nonzero anisotropy values at times long compared to the fluorescence lifetimes (r „). We now present evidence that another widely used microviscosity probe, pe-rylene, is also a hindered rotator when bound to lipid bilayers. In fact, it is the degree to which its rotations are hindered, and not its rotational rates, which determines the conventionally measured steady-state anisotropy. These observations illustrate the need for caution in the literal interpretation of membrane microviscosities derived from anisotropy measurements.
登录
查看更多内容
构建适用于细胞或组织水平生物样品的纳秒荧光系统
影响因子：
1.5
作者：
K. Kinosita;S. Mitaku;A. Ikegami;N. Ohbo;T. Kunii
通讯作者：
T. Kunii
大肠菌素 E1 引起大肠杆菌细胞包膜微粘度增加的证据。
影响因子：
2.9
作者：
S. Helgerson;W. Cramer;J. M. Harris;F. Lytle
通讯作者：
F. Lytle
磷脂分散体中荧光偏振的测量和解释。
DOI：
--
发表时间：
1976
期刊：
Biochimica et Biophysica Acta
影响因子：
--
作者：
C. Bashford;C. Morgan;G. Radda
通讯作者：
G. Radda
位于脂质双层膜内的荧光探针的旋转。
影响因子：
3.4
作者：
K. Jacobson;D. Wobschall
通讯作者：
D. Wobschall
氧猝灭条件下的荧光各向异性测量作为量化荧光团去偏振旋转的方法。
影响因子：
2.9
作者：
J. Lakowicz;F. Prendergast;D. Hogen
通讯作者：
D. Hogen