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Combining Cryogenic Infrared Spectroscopy with Selective Enzymatic Cleavage for Determining Glycan Primary Structure.
Analytical Chemistry ( IF 7.4 ) Pub Date : 2020-01-08 , DOI: 10.1021/acs.analchem.9b04776
Irina Dyukova 1 , Eduardo Carrascosa 1 , Robert P Pellegrinelli 1 , Thomas R Rizzo 1
Affiliation  

Given the biological relevance and intrinsic structural complexity of glycans, increasing efforts are being directed toward developing a general glycan database that includes information from different analytical methods. As recently demonstrated, cryogenic infrared (IR) spectroscopy is a promising technique for glycan analysis, as it provides unique vibrational fingerprints of specific glycan isomer ions. One of the main goals of a glycan database is the identification and detailed characterization of unknown species. In this work, we combine enzymatic digestion with cryogenic IR-spectroscopy and demonstrate how it can be used for glycan identification. We measured the IR-spectra of a series of cationic glycan standards of increasing complexity and compared them with spectra of the same species after enzymatic cleavage of larger glycans. We show that the cryogenic IR spectra of the cleaved glycans are highly structured and virtually identical to those of standards after both single and multiple cleavages. Our results suggest that the combination of these methods represents a potentially powerful and specific approach for the characterization of unknown glycans.

中文翻译:

结合低温红外光谱和选择性酶解确定聚糖的一级结构。

考虑到聚糖的生物学相关性和固有的结构复杂性,人们正在加大力度开发通用的聚糖数据库,其中包括来自不同分析方法的信息。正如最近证明的那样,低温红外(IR)光谱技术是糖类分析的一种有前途的技术,因为它提供了特定糖类异构体离子的独特振动指纹。聚糖数据库的主要目标之一是未知物种的鉴定和详细表征。在这项工作中,我们将酶消化与低温红外光谱结合起来,证明了其可用于聚糖鉴定。我们测量了一系列阳离子聚糖标准品的红外光谱,这些标准品的复杂性不断提高,并将它们与酶裂解较大聚糖后的同一物种的光谱进行比较。我们显示,裂解的聚糖的低温红外光谱具有高度结构化,并且与单次和多次裂解后的标准品的红外光谱几乎相同。我们的结果表明,这些方法的组合代表了未知糖聚糖表征的潜在强大且特定的方法。
更新日期:2020-01-08
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