Minimal Effects of Macromolecular Crowding on an Intrinsically Disordered Protein: A Small-Angle Neutron Scattering Study
Minimal Effects of Macromolecular Crowding on an Intrinsically Disordered Protein: A Small-Angle Neutron Scattering Study
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DOI：
10.1016/j.bpj.2013.12.003
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发表时间：
2014-02-18
影响因子：
3.4
通讯作者：
Argyle, Brian
中科院分区：
文献类型：
--
作者：
Goldenberg, David P.;Argyle, Brian
用小角中子散射研究了两种球状蛋白质，即牛胰蛋白酶抑制剂和马高铁肌红蛋白对一种内在无序的蛋白质--噬菌体Lambda的N蛋白--构象集合的影响。用H-2均匀标记lambda N蛋白，并调整样品中D2O的浓度，以匹配未标记的拥挤蛋白的中子散射对比度，从而掩盖它们对散射图谱的贡献。当样品中含有0.12克/毫升的牛胰蛋白酶抑制物或0.2克/毫升的高铁肌红蛋白时，记录了来自氚的波长N的散射。不拥挤的蛋白质的旋转半径估计为30埃，并被发现对人群的存在非常不敏感，变化幅度为
Small-angle neutron scattering was used to study the effects of macromolecular crowding by two globular proteins, i.e., bovine pancreatic trypsin inhibitor and equine metmyoglobin, on the conformational ensemble of an intrinsically disordered protein, the N protein of bacteriophage lambda. The lambda N protein was uniformly labeled with H-2, and the concentrations of D2O in the samples were adjusted to match the neutron scattering contrast of the unlabeled crowding proteins, thereby masking their contribution to the scattering profiles. Scattering from the deuterated lambda N was recorded for samples containing up to 0.12 g/mL bovine pancreatic trypsin inhibitor or 0.2 g/mL metmyoglobin. The radius of gyration of the uncrowded protein was estimated to be 30 angstrom and was found to be remarkably insensitive to the presence of crowders, varying by