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Nutrient-dependent structural changes in S. aureus peptidoglycan revealed by solid-state NMR spectroscopy.固相结合态 NMR 光谱揭示金黄色葡萄球菌肽聚糖的营养依赖性结构变化。
Biochemistry. 2012 Oct 16;51(41):8143-53. doi: 10.1021/bi3012115. Epub 2012 Oct 2.
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In-cell solid-state NMR as a tool to study proteins in large complexes.在细胞内固态 NMR 作为研究大复合物中蛋白质的工具。
Chembiochem. 2012 Mar 5;13(4):534-7. doi: 10.1002/cbic.201100721. Epub 2012 Feb 1.
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Revealing protein structures in solid-phase peptide synthesis by 13C solid-state NMR: evidence of excessive misfolding for Alzheimer's β.通过 13C 固态 NMR 揭示固相肽合成中的蛋白质结构：阿尔茨海默病 β 过度错误折叠的证据。
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4
Solid-state nuclear magnetic resonance (NMR) spectroscopy of human immunodeficiency virus gp41 protein that includes the fusion peptide: NMR detection of recombinant Fgp41 in inclusion bodies in whole bacterial cells and structural characterization of purified and membrane-associated Fgp41.固态核磁共振（NMR）光谱法研究人类免疫缺陷病毒 gp41 蛋白，包括融合肽：在全细菌细胞包涵体中用 NMR 检测重组 Fgp41 以及纯化和膜结合 Fgp41 的结构特征。
Biochemistry. 2011 Nov 22;50(46):10013-26. doi: 10.1021/bi201292e. Epub 2011 Oct 31.
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In situ structural characterization of a recombinant protein in native Escherichia coli membranes with solid-state magic-angle-spinning NMR.利用固态魔角旋转 NMR 对天然大肠杆菌膜中的重组蛋白进行原位结构表征。
J Am Chem Soc. 2011 Aug 17;133(32):12370-3. doi: 10.1021/ja204062v. Epub 2011 Jul 26.
6
The final conformation of the complete ectodomain of the HA2 subunit of influenza hemagglutinin can by itself drive low pH-dependent fusion.流感血凝素 HA2 亚基完整胞外结构域的最终构象本身可以驱动依赖 pH 值的低融合。
J Biol Chem. 2011 Apr 15;286(15):13226-34. doi: 10.1074/jbc.M110.181297. Epub 2011 Feb 3.
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19F NMR analysis of the antimicrobial peptide PGLa bound to native cell membranes from bacterial protoplasts and human erythrocytes.19F NMR 分析抗菌肽 PGLa 与细菌原生质体和人红细胞天然细胞膜的结合。
J Am Chem Soc. 2010 Jul 7;132(26):8822-4. doi: 10.1021/ja101608z.
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Solution structure of proinsulin: connecting domain flexibility and prohormone processing.胰岛素原的溶液结构：连接结构域的柔韧性与前激素加工。
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Comparative analysis of membrane-associated fusion peptide secondary structure and lipid mixing function of HIV gp41 constructs that model the early pre-hairpin intermediate and final hairpin conformations.比较分析模拟 HIV gp41 早期发夹前中间体和最终发夹构象的膜相关融合肽二级结构和脂质混合功能的构建体。
J Mol Biol. 2010 Mar 19;397(1):301-15. doi: 10.1016/j.jmb.2010.01.018. Epub 2010 Jan 18.
10
Hairpin folding of HIV gp41 abrogates lipid mixing function at physiologic pH and inhibits lipid mixing by exposed gp41 constructs.HIV gp41的发夹折叠在生理pH值下消除脂质混合功能，并通过暴露的gp41构建体抑制脂质混合。
Biochemistry. 2009 Mar 31;48(12):2714-22. doi: 10.1021/bi8019492.

通过固态核磁共振波谱技术对全细胞和细胞提取物中的重组蛋白进行定量分析。

Quantitation of recombinant protein in whole cells and cell extracts via solid-state NMR spectroscopy.

机构信息

Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, United States.

出版信息

Biochemistry. 2013 Jun 25;52(25):4285-7. doi: 10.1021/bi4007034. Epub 2013 Jun 17.

DOI:10.1021/bi4007034
PMID:23742073
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3734946/
Abstract

Recombinant proteins (RPs) are commonly expressed in bacteria followed by solubilization and chromatography. Purified RP yield can be diminished by losses at any step with very different changes in methods that can improve the yield. Time and labor can therefore be saved by first identifying the specific reason for the low yield. This study describes a new solid-state nuclear magnetic resonance approach to RP quantitation in whole cells or cell extracts without solubilization or purification. The method is straightforward and inexpensive and requires only ∼50 mL culture and a low-field spectrometer.

摘要

重组蛋白（RPs）通常在细菌中表达，然后进行溶解和色谱分离。纯化的 RP 产量可能会在任何步骤中损失，而不同的方法可以提高产量，从而导致产量变化非常大。因此，通过首先确定低产量的具体原因，可以节省时间和劳动力。本研究描述了一种新的固态核磁共振方法，无需溶解或纯化即可在全细胞或细胞提取物中定量 RP。该方法简单、廉价，仅需要约 50 mL 培养物和低场光谱仪。