Evaluation of immobilized polypeptides with different C‐terminal residues using argon gas‐cluster SIMS
Evaluation of immobilized polypeptides with different C‐terminal residues using argon gas‐cluster SIMS
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使用氩气簇 SIMS 评估具有不同 C 末端残基的固定化多肽
DOI：
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
K. Mochiji
中科院分区：
文献类型：
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作者：
S. Aoyagi;K. Moritani;K. Mochiji
气体团簇（GC）离子源，例如氩团簇源，由于其低能量射弹效应，已经被用于有机或生物材料的西姆斯中。最近，据报道发现西姆斯可对分子量为10000或更高的蛋白质进行完整离子检测。由于西姆斯能够控制每个原子的气体团簇大小和碰撞能量，因此被认为对于检测多肽单层中非常小的差异非常敏感，例如亮脑啡肽（H-Tyr-Gly-Gly-Phe-Leu-OH）和甲硫氨酸脑啡肽（H-Tyr-Gly-Gly-Phe-Met-OH）。在这项研究中，用西姆斯测量了两种脑啡肽，并比较了它们的光谱，以评估西姆斯对微小化学结构差异的灵敏度。通过将多肽（亮脑啡肽和甲硫氨酸）的N末端残基与戊二醛活化的氨基硅烷化ITO板结合，将其分别固定在铟和氧化锡（ITO）涂覆的玻璃板上。还制备不含多肽的氨基硅烷化ITO板作为对照样品。冷冻干燥后，用氩团簇离子源用TOF-西姆斯测量样品，然后分析TOF-西姆斯光谱以选择多肽特异性峰。结果，使用氩西姆斯检测到与没有重组的多肽相关的碎片离子。版权所有© 2010约翰威利父子有限公司.
Gas cluster (GC) ion sources, such as an argon cluster source, have been employed in SIMS of organic or biomaterials due to their low‐energy projectile effect. Recently, it was found that GC‐SIMS reportedly provides intact ion detection of proteins with molecular weights of 10000 or higher. Since GC‐SIMS enables control of the gas cluster size and collision energy per atom, it is considered sensitive for the detection of very small differences in the monolayers of polypeptides such as leu‐enkephalin (H‐Tyr‐Gly‐Gly‐Phe‐Leu‐OH) and met‐enkephalin (H‐Tyr‐Gly‐Gly‐Phe‐Met‐OH). In this study, both enkephalins were measured with GC‐SIMS and their spectra were compared to evaluate GC‐SIMS sensitivity to small chemical structure differences. The polypeptides, leu‐enkephalin and met‐enkephalin, were immobilized on indium‐ and tin oxide (ITO)‐coated glass plates, respectively, by binding their N terminal residues to glutaraldehyde‐activated aminosilanized‐ITO plates. An aminosilanized‐ITO plate without polypeptide was also prepared as the control sample. After freeze‐drying, the samples were measured with TOF‐SIMS with the argon cluster ion source, and then TOF‐SIMS spectra were analyzed to select peaks specific to the polypeptides. As a result, fragment ions related to the polypeptide without recombination were detected using the argon GC‐SIMS. Copyright © 2010 John Wiley & Sons, Ltd.