Peptide-metal ion interactions in solution: Detection by laser desorption time-of-flight mass spectrometry and electrospray ionization mass spectrometry†
Peptide-metal ion interactions in solution: Detection by laser desorption time-of-flight mass spectrometry and electrospray ionization mass spectrometry†
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溶液中肽-金属离子相互作用：通过激光解吸飞行时间质谱和电喷雾电离质谱检测†
DOI：
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
T. Yip
中科院分区：
文献类型：
--
作者：
T. Hutchens;R. Nelson;M. Allen;C. Li;T. Yip
采用基质辅助紫外激光解吸飞行时间质谱（LDTOF）和电喷雾质谱（ES）两种不同的软电离质谱方法研究了Cu（II）离子与溶液中金属结合肽的特异性相互作用。这些研究选择的金属结合肽是在人血浆金属运输蛋白富组氨酸糖蛋白表面发现的26个残基序列。在溶液中加入Cu（II）或Mn（II）离子前后，利用ES和LDTOF对（GHHPH） 5g肽进行了合成和评价。在没有添加金属离子的情况下，通过LDTOF和ES观察到肽的质量等于其计算质量（2903.0 Da）的0.5 Da以内。在Cu（II）的存在下，在大约63.9 Da （LDTOF）或62.3 Da （ES）的质量增量处观察到多达5个额外的峰；在相同的实验条件下，Mn没有与肽结合。通过LDTOF和ES，观察到的最大Cu结合容量（即5 g原子mol−1）表明每个（GHHPH）内部重复单元可以结合多达1个Cu。这种多肽-金属离子相互作用的化学计量学通过溶液中直接滴定法和固定化多肽的定量金属离子亲和色谱法进行了验证。因此，检测稳定的肽-金属配合物的能力似乎没有受到两种不同的挥发/电离方法的不同影响，这两种方法需要产生带电的完整分子离子。然而，LDTOF基体和ES溶液组成等实验条件会影响结合Cu原子的数量和化学计量。这些结果表明可以通过LDTOF和ES直接验证金属结合肽的液相结合能力。我们从这些研究中得出结论，其他金属-有机相互作用也可以通过这些快速灵敏的技术来研究。
The specific interaction of Cu(II) ions with metal-binding peptides in solution has been investigated by two different methods of soft ionization mass spectrometry, namely matrix-assisted ultraviolet laser desorption time-of-flight mass spectrometry (LDTOF) and electrospray ionization mass spectrometry (ES). The metal-binding peptide selected for these investigations is a 26-residue sequence found on the surface of the human plasma metal-transport protein histidine-rich glycoprotein. The peptide, (GHHPH)5 G, was synthesized and evaluated by ES and LDTOF before and after the addition of Cu(II) or Mn(II) ions in solution. In the absence of added metal ions, the peptide was observed to have a mass equal to within 0.5 Da of its calculated mass (2903.0 Da) by both LDTOF and ES. In the presence of Cu(II), up to five additional peaks were observed at mass increments of approximately 63.9 Da (LDTOF) or 62.3 Da (ES); Mn was not bound to the peptide under identical experimental conditions. By both LDTOF and ES, the maximum Cu-binding capacity observed (i.e., 5 g-atoms mol−1) demonstrated that up to 1 Cu could be bound per (GHHPH) internal repeat unit. This peptide-metal ion interaction stoichiometry was verified by direct titration in solution and, with immobilized peptide, by quantitative metal ion affinity chromatography. Thus, the ability to detect stable peptide-metal complexes did not appear to be differentially affected by the two different volatilization/ionization methods needed to generate charged intact molecular ions. The quantity and stoichiometry of bound Cu atoms was affected, however, by experimental conditions such as LDTOF matrix and ES solution composition. These results demonstrate the ability to verify directly the solution-phase binding capacity of metal-binding peptides by LDTOF and by ES. We conclude from these studies that other metallo-organic interactions may also be amenable to investigation by these rapid and sensitive techniques.
DOI：
10.1073/pnas.87.17.6873
发表时间：
1990-09-01
影响因子：
11.1
作者：
BEAVIS, RC;CHAIT, BT
通讯作者：
CHAIT, BT
DOI：
10.1002/rcm.1290031207
发表时间：
1989-12-01
期刊：
Rapid communications in mass spectrometry : RCM
影响因子：
--
作者：
Beavis, R C;Chait, B T
通讯作者：
Chait, B T
DOI：
10.1073/pnas.86.9.3094
发表时间：
1989-05-01
影响因子：
11.1
作者：
EISENSTEIN, E;MARKBY, DW;SCHACHMAN, HK
通讯作者：
SCHACHMAN, HK