Self-assembling peptide detergents stabilize isolated photosystem I on a dry surface for an extended time.
Self-assembling peptide detergents stabilize isolated photosystem I on a dry surface for an extended time.
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自组装肽洗涤剂会在干燥的表面上稳定孤立的光系统I。
DOI：
10.1371/journal.pbio.0030230
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发表时间：
2005-07
期刊：
影响因子：
9.8
通讯作者：
中科院分区：
文献类型：
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作者：
我们使用了一类设计的肽洗涤剂，以稳定光系统I（PS-I）后，延长干燥下N2的镀金-Ni-NTA玻璃表面。PS-I是一种含叶绿素的膜蛋白复合物，是铁氧还蛋白的主要还原剂和质体蓝素的电子受体。我们用化学去污剂从菠菜叶绿体类囊体中分离出了该复合物。与PS-I复合物相关的叶绿素分子在−196.15 °C下提供650至800 nm之间的固有稳态发射光谱，反映了色素-蛋白质相互作用的组织。在没有洗涤剂的情况下，荧光最大值从大约735 nm到大约685 nm的大的蓝移表明捕光亚基组织的破坏，从而揭示了叶绿素-蛋白质相互作用。常用的膜蛋白稳定去污剂N-十二烷基-β-D-麦芽糖苷和N-辛基-β-D-葡萄糖苷仅部分稳定了约735 nm的复合物，光谱位移约为685 nm。然而，在干燥之前，以增加的浓度添加肽去污剂乙酰基- AAAAAAK显著地稳定了PS-I复合物。此外，在乙酰基- AAAAAAK的存在下，PS-I复合物在室温下以干燥形式稳定至少3周。另一种肽去污剂，乙酰基-VVVVVD，也稳定了复合物，但程度较低。这些观察结果表明，肽去污剂可以有效地稳定膜蛋白在固态。这些设计的肽洗涤剂可以促进不同类型的膜蛋白的研究。 设计肽洗涤剂可以促进不同类型的膜蛋白的研究。
We used a class of designed peptide detergents to stabilize photosystem I (PS-I) upon extended drying under N2 on a gold-coated-Ni-NTA glass surface. PS-I is a chlorophyll-containing membrane protein complex that is the primary reducer of ferredoxin and the electron acceptor of plastocyanin. We isolated the complex from the thylakoids of spinach chloroplasts using a chemical detergent. The chlorophyll molecules associated with the PS-I complex provide an intrinsic steady-state emission spectrum between 650 and 800 nm at −196.15 °C that reflects the organization of the pigment-protein interactions. In the absence of detergents, a large blue shift of the fluorescence maxima from approximately 735 nm to approximately 685 nm indicates a disruption in light-harvesting subunit organization, thus revealing chlorophyll−protein interactions. The commonly used membrane protein-stabilizing detergents, N-dodecyl-β-D-maltoside and N-octyl-β-D-glucoside, only partially stabilized the approximately 735-nm complex with approximately 685-nm spectroscopic shift. However, prior to drying, addition of the peptide detergent acetyl- AAAAAAK at increasing concentration significantly stabilized the PS-I complex. Moreover, in the presence of acetyl- AAAAAAK, the PS-I complex is stable in a dried form at room temperature for at least 3 wk. Another peptide detergent, acetyl-VVVVVVD, also stabilized the complex but to a lesser extent. These observations suggest that the peptide detergents may effectively stabilize membrane proteins in the solid-state. These designed peptide detergents may facilitate the study of diverse types of membrane proteins. A designer peptide detergent may facilitate the study of diverse types of membrane proteins.