De novo sequencing of novel neuropeptides directly from Ascaris suum tissue using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight.
De novo sequencing of novel neuropeptides directly from Ascaris suum tissue using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight.
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使用基质辅助激光解吸/电离飞行时间/飞行时间直接从猪蛔虫组织中对新型神经肽进行从头测序。
DOI：
10.1002/rcm.1240
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Stretton,AntonyO
中科院分区：
文献类型：
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作者：
Yew,JoanneY;Dikler,Sergei;Stretton,AntonyO
通过基质辅助激光解吸/电离质谱 (MALDI-MS) 对组织进行直接分析，可以快速分析生物分子，同时最大限度地减少样品损失或降解，并降低化学修饰的可能性。然而，由于组织的复杂性和单个细胞中内源肽的数量较少，仍然存在相当大的挑战需要克服。由于单个神经元尺寸小且每个细胞的肽很少，这些问题在猪蛔虫中更加严重。为了解决这些困难，最近开发的基质辅助激光解吸/电离飞行时间/飞行时间（MALDI-TOF/TOF）技术与靶向衍生化相结合，以便直接对来自蛔虫神经组织的新型神经肽进行测序。对蛔虫组织的直接 MALDI-TOF/TOF 分析提供了先前表征的神经肽以及在其他线虫中发现的具有同源物的三种新肽的完整氨基酸序列。这些结果证明了一种使用 MALDI-TOF/TOF 直接从组织中快速表征亚飞摩尔量肽的方法。版权所有 © 2003 约翰·威利父子有限公司
Direct analysis of tissue by matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) allows for the rapid profiling of biological molecules with minimal loss of sample or degradation and reduced likelihood of chemical modification. However, there are still considerable challenges to overcome due to the complexity of tissue and the low quantity of endogenous peptide in a single cell. These problems are exacerbated in the nematodeAscaris suumbecause of the small size of individual neurons and the paucity of peptide per cell. In an effort to address these difficulties, the recently developed matrix‐assisted laser desorption/ionization time‐of‐flight/time‐of‐flight (MALDI‐TOF/TOF) technology was used in combination with an on‐target derivatization in order to sequence novel neuropeptides directly fromAscarisnervous tissue. Direct MALDI‐TOF/TOF analysis ofAscaristissue provided the complete amino acid sequences for a previously characterized neuropeptide as well as for three novel peptides with homologues found in other nematodes. These results demonstrate a method for the rapid characterization of sub‐femtomolar amounts of peptide directly from tissue using MALDI‐TOF/TOF. Copyright © 2003 John Wiley & Sons, Ltd.