Incorporation of cytochrome oxidase into cardiolipin bilayers and induction of nonlamellar phases.

Incorporation of cytochrome oxidase into cardiolipin bilayers and induction of nonlamellar phases.
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将细胞色素氧化酶掺入心磷脂双层并诱导非层状相。

DOI:
10.1021/bi00473a018
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发表时间:
1990
期刊:
影响因子:
2.9
通讯作者:
Marsh,D
Marsh,D
中科院分区:
生物学3区
文献类型:
--
作者:
Powell,GL;Knowles,PF;Marsh,D

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材料与方法牛心脏心磷脂钠盐和细胞色素c(VI级)均来自Sigma化学公司。(St.路易斯,密苏里州)。二肉豆蔻酰磷脂酰胆碱纯化物来自Fluka(布克斯,瑞士)。自旋标记的马来酰亚胺(4-马来酰亚氨基-2,2,6,6-四甲基-1-哌啶氧基)来自Syva(加利福尼亚州帕洛阿尔托)。采用俞等的方法从牛心中提取细胞色素氧化酶。(1975),并在Cable和Powell(1980)中进行了修改。根据Knowles等人描述的方法进行二肉豆蔻酰磷脂酰胆碱的脂质替代。(1979),但是从缓冲液中省略蔗糖和KCl。进一步的细节在Powellet等人中给出。(1987年)。通过类似的方法进行心磷脂的脂质替代,不同之处在于使用10倍过量的心磷脂进行两次交换。心磷脂样品在N2下处理以抑制脂质过氧化。将最终的心磷脂替代制剂通过1、2或4 M NaCl离心,以从未掺入的脂质中分离蛋白质/脂质复合物。然后蔗糖密度梯度离心证明复合物沉淀为单一条带并且不含游离脂质。缩写:NMR,核磁共振; ESR,电子自旋共振; STESR,饱和转移ESR; V/,相对于场调制检测到90异相的二次谐波ESR吸收信号; EM,电子显微镜; TLC,薄层色谱法; Ln,流体层状脂质相; Hn,倒置六边形脂质相。
Materials and MethodsBeef heart cardiolipin (sodium salt) and cytochrome c (grade VI) were obtained from Sigma Chemical Co.(St. Louis, MO). Dimyristoylphosphatidylcholine puriss was from Fluka (Buchs, Switzerland). Spin-labeled maleimide (4-maleimido-2, 2, 6, 6-tetramethyl-l-piperidineoxyl) was from Syva (Palo Alto, CA). Cytochrome oxidase was prepared from beef heart by using the method of Yu et al.(1975) with modifications given in Cable and Powell (1980). Lipid substitution by di-myristoylphosphatidylcholine was carried out according to the method described by Knowles et al.(1979), butwith omission of sucrose and KC1 from the buffers. Further details are given in Powellet al.(1987). Lipid substitution by cardiolipin was performed by a similar method, except that two exchanges with a 10-fold excess of cardiolipin were employed. Cardiolipin samples were handled under N2 to inhibit lipid peroxidation. The final cardiolipin-substituted preparations were centrifuged through either 1, 2, or 4 M NaCl to resolve the protein/lipid complexes from unincorporated lipid. Sucrose density gradient centrifugation then demonstrated that the complexes sedimented as a single band and contained no free lipid. Salt concentration was varied by resuspending and washing the1 Abbreviations: NMR, nuclear magnetic resonance; ESR, electron spin resonance; STESR, saturation-transfer ESR; V/, second harmonic ESR absorption signal detected 90 out-of-phase with respect to the field modulation; EM, electron microscopy; TLC, thin-layer chromatography; L „, fluid lamellar lipid phase; Hn, inverted hexagonal lipid phase.
DOI: 10.1016/s0021-9258(19)41825-5
发表时间: 1975
期刊: The Journal of biological chemistry
影响因子: --
作者:
C. Yu;L. Yu;T. King
通讯作者: T. King
DOI: 10.1016/0022-2364(84)90295-6
发表时间: 1984
影响因子: 2.2
作者:
M. A. Hemminga;P. A. D. Jager;D. Marsh;P. Fajer
通讯作者: P. Fajer
分离的光感受器膜脂的相行为由二价阳离子调节
DOI: 10.1016/0014-5793(84)80329-4
发表时间: 1984
期刊: FEBS Letters
影响因子: 3.5
作者:
L. Mollevanger;W. D. de Grip
通讯作者: W. D. de Grip
蛋白质-脂质相互作用的进展,第 2 卷
DOI: 10.1016/0014-5793(86)81366-7
发表时间: 1986
期刊: FEBS Letters
影响因子: 3.5
作者:
G. Roberts
通讯作者: G. Roberts
DOI: 10.1016/0005-2736(85)90409-2
发表时间: 1985
期刊: Biochimica et biophysica acta
影响因子: --
作者:
G. Powell;P. Knowles;D. Marsh
通讯作者: D. Marsh