一种用于蛋白质稳定性的定量、高通量筛选方法。
A quantitative, high-throughput screen for protein stability.
作者信息
Ghaemmaghami S, Fitzgerald M C, Oas T G
机构信息
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
出版信息
Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8296-301. doi: 10.1073/pnas.140111397.
Abstract
In proteomic research, it is often necessary to screen a large number of polypeptides for the presence of stable structure. Described here is a technique (referred to as SUPREX, stability of unpurified proteins from rates of H/D exchange) for measuring the stability of proteins in a rapid, high-throughput fashion. The method uses hydrogen exchange to estimate the stability of microgram quantities of unpurified protein extracts by using matrix-assisted laser desorption/ionization MS. The stabilities of maltose binding protein and monomeric lambda repressor variants determined by SUPREX agree well with stability data obtained from conventional CD denaturation of purified protein. The method also can detect the change in stability caused by the binding of maltose to maltose binding protein. The results demonstrate the precision of the method over a wide range of stabilities.
摘要
在蛋白质组学研究中，常常需要筛选大量多肽以确定其是否存在稳定结构。本文介绍了一种技术（称为SUPREX，即通过氢/氘交换速率测定未纯化蛋白质的稳定性），可快速、高通量地测量蛋白质的稳定性。该方法利用氢交换，通过基质辅助激光解吸/电离质谱来估算微克量未纯化蛋白质提取物的稳定性。通过SUPREX测定的麦芽糖结合蛋白和单体λ阻遏物变体的稳定性，与通过传统的纯化蛋白质圆二色性变性获得的稳定性数据高度吻合。该方法还能检测麦芽糖与麦芽糖结合蛋白结合所引起的稳定性变化。结果证明了该方法在广泛的稳定性范围内的精确性。
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本文引用的文献
1
Rate and equilibrium constants for protein unfolding and refolding determined by hydrogen exchange-mass spectrometry.通过氢交换质谱法测定的蛋白质展开和重折叠的速率及平衡常数。
Anal Biochem. 1999 Dec 15;276(2):150-60. doi: 10.1006/abio.1999.4347.
2
Protein conformational stabilities can be determined from hydrogen exchange rates.蛋白质构象稳定性可通过氢交换速率来确定。
Nat Struct Biol. 1999 Oct;6(10):910-2. doi: 10.1038/13273.
3
Protein misfolding, evolution and disease.蛋白质错误折叠、进化与疾病。
Trends Biochem Sci. 1999 Sep;24(9):329-32. doi: 10.1016/s0968-0004(99)01445-0.
4
Denaturant mediated unfolding of both native and molten globule states of maltose binding protein are accompanied by large deltaCp's.变性剂介导的麦芽糖结合蛋白天然态和熔球态的去折叠都伴随着较大的等压热容变化。
Protein Sci. 1999 Aug;8(8):1689-95. doi: 10.1110/ps.8.8.1689.
5
Proteomics: quantitative and physical mapping of cellular proteins.蛋白质组学：细胞蛋白质的定量与物理图谱分析
Trends Biotechnol. 1999 Mar;17(3):121-7. doi: 10.1016/s0167-7799(98)01245-1.
6
Measurement of amide hydrogen exchange by MALDI-TOF mass spectrometry.通过基质辅助激光解吸电离飞行时间质谱法测量酰胺氢交换
Anal Chem. 1998 Oct 1;70(19):3987-95. doi: 10.1021/ac980553g.
7
Folding kinetics of a fluorescent variant of monomeric lambda repressor.单体λ阻遏物荧光变体的折叠动力学
Biochemistry. 1998 Jun 23;37(25):9179-85. doi: 10.1021/bi980356b.
8
Characterization of the free energy spectrum of peptostreptococcal protein L.消化链球菌蛋白L自由能谱的表征
Fold Des. 1997;2(5):271-80. doi: 10.1016/S1359-0278(97)00038-2.
9
Strategies for the in vitro evolution of protein function: enzyme evolution by random recombination of improved sequences.蛋白质功能的体外进化策略：通过改进序列的随机重组进行酶进化。
J Mol Biol. 1997 Sep 26;272(3):336-47. doi: 10.1006/jmbi.1997.1252.
10
The energy landscape of a fast-folding protein mapped by Ala-->Gly substitutions.通过丙氨酸到甘氨酸取代绘制的快速折叠蛋白质的能量景观。
Nat Struct Biol. 1997 Apr;4(4):305-10. doi: 10.1038/nsb0497-305.
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