Effect of lipid membrane structure on the adenosine 5'-triphosphate hydrolyzing activity of the calcium-stimulated adenosinetriphosphatase of sarcoplasmic reticulum.
Effect of lipid membrane structure on the adenosine 5'-triphosphate hydrolyzing activity of the calcium-stimulated adenosinetriphosphatase of sarcoplasmic reticulum.
复制标题
脂质膜结构对钙刺激肌浆网腺苷三磷酸酶5-三磷酸腺苷水解活性的影响。
DOI：
10.1021/bi00527a011
复制
发表时间：
1981
期刊：
影响因子：
2.9
通讯作者：
Meissner,G
中科院分区：
文献类型：
--
作者：
Moore,BM;Lentz,BR;Hoechli,M;Meissner,G
Bryant M. Moore, 1 Barry R. Lentz， Mathias Hoechli和Gerhard Meissner*摘要：从兔骨骼肌肌浆网膜中分离出一种活性的Ca2+刺激的，Mg2+依赖的腺苷三磷酸酶（Ca2+- atp酶），通过一种新开发的脂质替代程序，已被纳入到二脲酰-，二肉豆醇-，二戊烷酰-，双棕榈酰-和棕榈酰磷脂酰胆碱双分子层中，取代了99%以上的内源性脂质。冷冻断口电镜显示尺寸均匀的膜状囊泡和对称分布的断口颗粒。利用二苯基六三烯荧光各向异性来定义重组膜相行为，并揭示了膜中的多个转变。酶分析表明，饱和磷脂酰基链在脂质替代膜的主要脂质相变温度（Tm）以下抑制了总atp酶活性和Ca2+依赖性磷酸酶的形成。在高于Tm的温度下，atp酶活性而非磷酸酶的形成严重依赖于酰基链的长度和双层厚度。二脲酰磷脂酰胆碱双分子层未见atp酶活性。使用非离子洗涤剂十二环二氧乙烯乙二醇单醚证明了活性的缺乏不是由于酶的不可逆失活。双分子层厚度的增加导致活性水平的增加。当双棕榈酰磷脂酰胆碱中的一种饱和脂肪酸被油酸取代时，活性又增加了2倍，油酸的酰基链长度与棕榈酸相当。这些结果表明，Ca2+- atp酶需要具有最小双层厚度和含有不饱和磷脂酰基链的“流体”膜才能发挥最佳功能。e肌浆网（SR）的Ca2+刺激，Mg2+依赖的atp酶（Ca2+-AT-Pase） 1通过atp依赖的肌原纤维空间钙的摄取来控制肌肉松弛（Tada等，1978）。Ca2+- atp酶是SR的主要组成部分，约占总膜蛋白的90% （Meissner, 1975）。流体，疏水的环境被认为是充分的酶活性所必需的。早期的研究表明，虽然充分的Ca2+- atp酶活性需要磷脂，但磷酸酶中间体的形成并不需要磷脂（Martonosi, 1969; Meissner & Fleischer, 1972）。最近，Dean和Tanford（1978）表明，洗涤剂溶解的Ca2+-ATP酶在每mol酶只存在1-3 mol磷脂的情况下可以水解ATP。为了确定磷脂酰基链结构在Ca2+- atp酶功能中的作用，该酶先前已被纳入由二酰基、双棕榈酰或二酰基磷脂酰胆碱组成的双层膜中（Hesketh等人，1976；Hidalgo等人，1976；Nakamura等人，1976）。对这些脂质替代酶制剂的研究得出了一个普遍的结论，即具有有序酰基链结构的磷脂抑制atp酶的活性。在熔融温度（T^ J）以上发生的无序酰基链构型支持atp酶活性。“刚性”磷脂酰基链环境对
Bryant M. Moore, 1 Barry R. Lentz, Mathias Hoechli, and Gerhard Meissner* abstract: An active Ca2+-stimulated, Mg2+-dependent adenosinetriphosphatase (Ca2+-ATPase) isolated fromrabbit skeletal muscle sarcoplasmic reticulum membranes has been incorporated into dilauroyl-, dimyristoyl-, dipentadecanoyl-, dipalmitoyl-, and palmitoyloleoylphosphatidylcholine bilayers by using a newly developed lipid-substitution procedure that replaces greater than 99% of the endogenous lipid. Freezefracture electron microscopy showed membranous vesicles of homogeneous size with symmetrically disposed fracture-face particles. Diphenylhexatriene fluorescence anisotropy was used to define the recombinant membranephase behavior and re-vealed more than one transition in themembranes. Enzymatic analysis indicated that saturated phospholipid acyl chains inhibited both overall ATPase activity and Ca2+-dependent phosphoenzyme formation below the main lipid phase transition temperature (Tm) of the lipid-replaced membranes. At temperatures above Tm, ATPase activity butnot phosphoenzyme formation was critically dependent on acyl chain length and thus bilayer thickness. No ATPase activity was observed in dilauroylphosphatidylcholine bilayers. Use of the nonionic detergent dodecyloctaoxyethylene glycol monoether demonstrated that the absence of activity was not due to irreversible inactivation of the enzyme. Increased bilayer thickness re-sulted in increased levels of activity. An additional 2-fold rise in activity was observed when one of the saturated fatty acids in dipalmitoylphosphatidylcholine was replaced by oleic acid, whose acyl chain has a fully extended length comparable to that of palmitic acid. These results indicate that the Ca2+-ATPase requires for optimal function a “fluid” membrane with a minimal bilayer thickness and containing unsaturated phospholipid acyl chains. e Ca2+-stimulated, Mg2+-dependent ATPase (Ca2+-AT-Pase) 1 of sarcoplasmic reticulum (SR) controls muscle re-laxation through ATP-dependent uptake of calcium from the muscle myofibrillar space (Tada et al., 1978). The Ca2+-ATPase is a major component of SR, accounting for about 90% of the total membrane protein (Meissner, 1975). A fluid, hydrophobic environment is believed to be required for full enzymatic activity. Early studies indicated that while full Ca2+-ATPase activity required phospholipid, formation of the phosphoenzyme intermediate did not (Martonosi, 1969; Meissner & Fleischer, 1972). Recently, Dean & Tanford (1978) showed that the detergent-solubilized Ca2+-ATPase can hydrolyze ATP in thepresence of only 1-3 mol of phos-pholipid per mol of enzyme. In orderto determine the role of phospholipid acyl chain structure in Ca2+-ATPase function, the enzyme has been incorporated previously into bilayer membranes composed of dioleoyl-, dipalmitoyl-, or dimyristoylphosphatidylcholine (Hesketh et al., 1976; Hidalgo et al., 1976; Nakamura et al., 1976). Studies with these lipid-replaced enzyme preparations led to the general conclusion that phospholipids with an or-dered acyl chain configuration inhibited ATPase activity. The disordered acyl chain configuration occurring above the melting temperature (T^ J supported ATPase activity. The effect of a “rigid” phospholipid acyl chain environment on
登录
查看更多内容
重建肌浆网囊泡的渗透性。
DOI：
10.1016/0005-2736(81)90466-1
发表时间：
1981
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Young,RC;Allen,R;Meissner,G
通讯作者：
Meissner,G
DOI：
10.1016/s0021-9258(19)69855-8
发表时间：
1981-02
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
A. Johannsson;C. Keightley;G. Smith;C. D. Richards;T. Hesketh;J. Metcalfe
通讯作者：
A. Johannsson;C. Keightley;G. Smith;C. D. Richards;T. Hesketh;J. Metcalfe
DOI：
10.1016/0005-2736(80)90031-0
发表时间：
1980-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
GOMEZFERNANDEZ, JC;GONI, FM;CHAPMAN, D
通讯作者：
CHAPMAN, D
ATP 依赖性钙泵的磷脂头基特异性。
DOI：
10.1016/0005-2736(78)90201-8
发表时间：
1978
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
J. P. Bennett;Gerry A. Smith;M. Houslay;T. Hesketh;J. Metcalfe;G. B. Warren
通讯作者：
G. B. Warren
影响因子：
4.8
作者：
A. Martonosi
通讯作者：
A. Martonosi