一种使用电喷雾电离质谱法测定肽和蛋白质中碱性氨基酸残基数目的简单而稳健的方法。
A simple and robust method for determining the number of basic sites in peptides and proteins using electrospray ionization mass spectrometry.
机构信息
Department of Chemistry, University of California, Berkeley, California 94720-1460, United States.
出版信息
Anal Chem. 2011 Mar 15;83(6):2210-4. doi: 10.1021/ac1031012. Epub 2011 Feb 21.
Abstract
A solution additive has been discovered that can be used to measure the number of basic sites in a peptide or protein using electrospray ionization (ESI) mass spectrometry. Addition of millimolar amounts of perchloric acid (HClO(4)) to aqueous solutions that contain peptides or proteins results in the noncovalent adduction of HClO(4) molecules to the multiply charged ions formed by ESI. For 18 oligopeptides and proteins, ranging in molecular weight from 0.5 to 18.3 kDa, the sum of the number of protons plus maximum number of HClO(4) molecules adducted to the lower charge state ions is equal to the number of basic sites in the molecule. This method provides a rapid means of obtaining information about the composition of a peptide or protein and does not require high-resolution measurements or any instrumental or experimental modifications.
摘要
现已发现一种溶液添加剂，可用于使用电喷雾电离 (ESI) 质谱法测定肽或蛋白质中的碱性基团数量。在含有肽或蛋白质的水溶液中添加毫摩尔浓度的高氯酸 (HClO(4)) 会导致 HClO(4)分子非共价地加合到 ESI 形成的多电荷离子上。对于分子量在 0.5 至 18.3 kDa 之间的 18 个寡肽和蛋白质，质子数加上与低电荷状态离子加合的最大 HClO(4)分子数的总和等于分子中的碱性基团数量。该方法提供了一种快速获取肽或蛋白质组成信息的方法，不需要高分辨率测量或任何仪器或实验修改。
相似文献
1
A simple and robust method for determining the number of basic sites in peptides and proteins using electrospray ionization mass spectrometry.一种使用电喷雾电离质谱法测定肽和蛋白质中碱性氨基酸残基数目的简单而稳健的方法。
Anal Chem. 2011 Mar 15;83(6):2210-4. doi: 10.1021/ac1031012. Epub 2011 Feb 21.
2
Charging of Proteins in Native Mass Spectrometry.天然质谱法中的蛋白质荷电。
J Am Soc Mass Spectrom. 2017 Feb;28(2):332-340. doi: 10.1007/s13361-016-1517-7. Epub 2016 Oct 12.
3
Effects of select anions from the Hofmeister series on the gas-phase conformations of protein ions measured with traveling-wave ion mobility spectrometry/mass spectrometry.手性离子淌度谱/质谱法测定荷质比相同的蛋白质离子在不同溶剂中的气相构象
J Am Soc Mass Spectrom. 2011 Nov;22(11):1978-90. doi: 10.1007/s13361-011-0238-1. Epub 2011 Sep 13.
4
Effects of anions on the positive ion electrospray ionization mass spectra of peptides and proteins.阴离子对肽和蛋白质正离子电喷雾电离质谱的影响。
Anal Chem. 1994 Sep 15;66(18):2898-904. doi: 10.1021/ac00090a017.
5
Supercharged protein and peptide ions formed by electrospray ionization.通过电喷雾电离形成的增压蛋白质和肽离子。
Anal Chem. 2001 Apr 1;73(7):1455-60. doi: 10.1021/ac001251t.
6
Reactive-electrospray-assisted laser desorption/ionization for characterization of peptides and proteins.用于肽和蛋白质表征的反应性电喷雾辅助激光解吸/电离
Anal Chem. 2008 Sep 15;80(18):6995-7003. doi: 10.1021/ac800870c. Epub 2008 Aug 7.
7
Anion effects on sodium ion and acid molecule adduction to protein ions in electrospray ionization mass spectrometry.电喷雾电离质谱中负离子对钠离子和酸分子与蛋白质离子加合的影响。
J Am Soc Mass Spectrom. 2011 Nov;22(11):1968-77. doi: 10.1007/s13361-011-0218-5. Epub 2011 Aug 9.
8
Multiply charged negative ions by electrospray ionization of polypeptides and proteins.通过多肽和蛋白质的电喷雾电离产生多电荷负离子。
Anal Chem. 1992 Jan 1;64(1):81-8. doi: 10.1021/ac00025a015.
9
Effects of cations on protein and peptide charging in electrospray ionization from aqueous solutions.阳离子对水溶液电喷雾电离中蛋白质和肽带电情况的影响。
J Am Soc Mass Spectrom. 2014 Jun;25(6):918-27. doi: 10.1007/s13361-014-0864-5. Epub 2014 Apr 12.
10
Charge-reduced nano electrospray ionization combined with differential mobility analysis of peptides, proteins, glycoproteins, noncovalent protein complexes and viruses.电荷降低的纳米电喷雾电离结合肽、蛋白质、糖蛋白、非共价蛋白质复合物和病毒的差分迁移率分析
J Mass Spectrom. 2001 Sep;36(9):1038-52. doi: 10.1002/jms.208.
引用本文的文献
1
Direct ESI-MS of Ionic Liquids Using High-Pressure Electrospray From Large-Bore Emitters Made of Micropipette Tips.使用由微量移液器尖端制成的大口径发射器进行高压电喷雾对离子液体进行直接电喷雾电离质谱分析。
Mass Spectrom (Tokyo). 2024;13(1):A0148. doi: 10.5702/massspectrometry.A0148. Epub 2024 Jul 17.
2
Top-down mass spectrometry of intact phosphorylated β-casein: Correlation between the precursor charge state and internal fragments.完整磷酸化β-酪蛋白的自上而下质谱分析：前体电荷状态与内部片段之间的相关性。
J Mass Spectrom. 2019 Jun;54(6):527-539. doi: 10.1002/jms.4364.
3
Flame Atmospheric Pressure Chemical Ionization Coupled with Negative Electrospray Ionization Mass Spectrometry for Ion Molecule Reactions.火焰常压化学电离耦合负电喷雾电离质谱用于离子分子反应。
J Am Soc Mass Spectrom. 2017 Jul;28(7):1473-1481. doi: 10.1007/s13361-017-1688-x. Epub 2017 May 15.
4
Effective protein separation by coupling hydrophobic interaction and reverse phase chromatography for top-down proteomics.通过疏水相互作用与反相色谱联用实现自上而下蛋白质组学中的有效蛋白质分离。
Anal Chem. 2014 Aug 5;86(15):7899-906. doi: 10.1021/ac501836k. Epub 2014 Jul 9.
5
Insights into the mechanism of protein electrospray ionization from salt adduction measurements.通过盐加成测量深入了解蛋白质电喷雾电离机制。
J Am Soc Mass Spectrom. 2014 Aug;25(8):1322-31. doi: 10.1007/s13361-014-0905-0. Epub 2014 May 17.
6
Cation-induced stabilization of protein complexes in the gas phase: mechanistic insights from hemoglobin dissociation studies.在气相中，阳离子诱导稳定蛋白质复合物：来自血红蛋白解离研究的机理见解。
J Am Soc Mass Spectrom. 2014 Apr;25(4):595-603. doi: 10.1007/s13361-013-0814-7. Epub 2014 Jan 23.
7
Dramatically stabilizing multiprotein complex structure in the absence of bulk water using tuned Hofmeister salts.使用调谐的霍夫迈斯特盐在没有大量水的情况下显著稳定多蛋白复合物结构。
Faraday Discuss. 2013;160:371-88; discussion 389-403. doi: 10.1039/c2fd20099f.
8
Solution additives that desalt protein ions in native mass spectrometry.用于在天然质谱中脱盐蛋白质离子的溶液添加剂。
Anal Chem. 2012 Sep 4;84(17):7511-7. doi: 10.1021/ac301629s. Epub 2012 Aug 21.
9
Bound cations significantly stabilize the structure of multiprotein complexes in the gas phase.结合阳离子能显著稳定气相中多蛋白复合物的结构。
Angew Chem Int Ed Engl. 2012 Jun 4;51(23):5692-5. doi: 10.1002/anie.201109127. Epub 2012 Apr 23.
10
Ion mobility-mass spectrometry for structural proteomics.离子淌度-质谱用于结构蛋白质组学。
Expert Rev Proteomics. 2012;9(1):47-58. doi: 10.1586/epr.11.75.
本文引用的文献
1
Solution-phase chelators for suppressing nonspecific protein-metal interactions in electrospray mass spectrometry.用于抑制电喷雾质谱中非特异性蛋白质 - 金属相互作用的溶液相螯合剂。
Anal Chem. 2009 Jun 15;81(12):5008-15. doi: 10.1021/ac900423x.
2
Protein-metal interactions of calmodulin and alpha-synuclein monitored by selective noncovalent adduct protein probing mass spectrometry.通过选择性非共价加合物蛋白质探测质谱法监测钙调蛋白与α-突触核蛋白的蛋白质-金属相互作用。
J Am Soc Mass Spectrom. 2008 Nov;19(11):1663-72. doi: 10.1016/j.jasms.2008.07.006. Epub 2008 Jul 16.
3
Using ESI-MS to probe protein structure by site-specific noncovalent attachment of 18-crown-6.利用电喷雾电离质谱通过18-冠-6的位点特异性非共价连接来探测蛋白质结构。
J Am Soc Mass Spectrom. 2006 Sep;17(9):1209-15. doi: 10.1016/j.jasms.2006.05.007. Epub 2006 Jun 12.
4
Mass defect labeling of cysteine for improving peptide assignment in shotgun proteomic analyses.用于改进鸟枪法蛋白质组分析中肽段鉴定的半胱氨酸质量缺陷标记
Anal Chem. 2006 May 15;78(10):3417-23. doi: 10.1021/ac0600407.
5
Pulsed dual electrospray ionization for ion/ion reactions.用于离子/离子反应的脉冲双电喷雾电离
J Am Soc Mass Spectrom. 2005 Nov;16(11):1750-6. doi: 10.1016/j.jasms.2005.07.013. Epub 2005 Sep 22.
6
Buffer loading for counteracting metal salt-induced signal suppression in electrospray ionization.用于抵消电喷雾电离中金属盐诱导信号抑制的缓冲液加载
Anal Chem. 2004 Jul 15;76(14):3944-50. doi: 10.1021/ac049724+.
7
Selective molecular recognition of arginine by anionic salt bridge formation with bis-phosphate crown ethers: implications for gas phase peptide acidity from adduct dissociation.通过与双磷酸冠醚形成阴离子盐桥对精氨酸进行选择性分子识别：加合物解离对气相肽酸度的影响。
J Am Soc Mass Spectrom. 2004 Apr;15(4):616-24. doi: 10.1016/j.jasms.2003.12.015.
8
Nanoelectrospray ionization of protein mixtures: solution pH and protein pI.蛋白质混合物的纳米电喷雾电离：溶液pH值与蛋白质等电点
Anal Chem. 2004 Feb 15;76(4):1165-74. doi: 10.1021/ac035209k.
9
Probing of arginine residues in peptides and proteins using selective tagging and electrospray ionization mass spectrometry.利用选择性标记和电喷雾电离质谱法探测肽和蛋白质中的精氨酸残基。
J Mass Spectrom. 2003 Aug;38(8):891-9. doi: 10.1002/jms.477.
10
Mechanism of charging and supercharging molecules in electrospray ionization.电喷雾电离中分子充电和增压的机制。
J Am Chem Soc. 2003 Feb 26;125(8):2319-27. doi: 10.1021/ja021202t.
Zotero 插件