Dynamics of a monomeric insulin analogue: testing the molten-globule hypothesis.
Dynamics of a monomeric insulin analogue: testing the molten-globule hypothesis.
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单体胰岛素类似物的动力学：检验熔球假说。
DOI：
10.1021/bi00057a006
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发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
Weiss,MA
中科院分区：
文献类型：
--
作者：
Hua,QX;Ladbury,JE;Weiss,MA
材料和方法SDPI由Eli Lilly & Co.（印第安纳州印第安纳波利斯）。通过280 nm处的吸光度测量蛋白质浓度。1-苯胺基-8-萘磺酸盐购自Molecular Probes，Inc.（尤金，OR）。使用Shimadzu分光荧光光度计（型号RF 5000 U）获得荧光光谱;使用1-cm路径长度的石英比色皿。使用配备有温度控制单元的Aviv分光光度计获得CD光谱;使用1-mm路径长度的石英比色皿。在哈佛医学院NMR设备和MIT弗朗西斯比特国家磁铁实验室在500 MHz下获得1H-NMR谱。使用核黄素（4 mM）作为三重态探针进行光化学动态核极化（photo-CIDNP）研究;光源是488 nm的Innova-6连续氩激光器。如所述计算光-暗差光谱（韦斯等人，1989年）。1H-NMR化学位移参考乙酸的甲基共振（假定为2.06ppm）。在H2O或20%氘代乙酸-80%H2O中获得的光谱中，溶剂共振通过预饱和而衰减。差示扫描量热法（DSC）实验在DASM-4微量热计（Biopribor，Puschino，Russia）中在2巴的过压下以1
MATERIALS AND METHODSDPI was provided by Eli Lilly & Co.(Indianapolis, IN). Protein concentration was measured by absorbance at 280 nm. l-Anilino-8-naphthalenesulfonate was purchased from Molecular Probes, Inc.(Eugene, OR). Fluorescence spectra were obtained using a Shimadzu spectrofluorophotometer (Model RF5000U); a 1-cm path-length quartz cuvette was used. CD spectra were obtained using an Aviv spectropo-larimeter equipped with a temperature-control unit; a 1-mm path-length qua rtz cuvette was used. 1 H-NMR spectra were obtained at 500 MHz at the Harvard Medical School NMR Facility and MIT Francis Bitter National Magnet Laboratory. Photochemical dynamic nuclear polarization (photo-CIDNP) studies were performed using riboflavin (4 mM) as triplet-state probe; the light source was an Innova-6 continuous argon laser at 488 nm. Light-minus-dark difference spectra were calculated as described (Weiss et al., 1989).'H-NMR chemical shifts were referenced to the methyl resonance of acetic acid (assumed to be 2.06 ppm). In spectra obtained in H20 or 20% deuterated acetic acid-80% H20 the solvent resonance was attenuated by presaturation. Differential scanning calorimetry (DSC) experiments were performed in a DASM-4 microcalorimeter (Biopribor, Puschino, Russia) under an excess pressure of 2 bar and with a heating rate of 1