PDF(Pubmed)
Abstract：
A certified reference material of ricin (CRM-LS-1) was produced by the EuroBioTox consortium to standardise the analysis of this biotoxin. This study established the N-glycan structures and proportions including their loci and occupancy of ricin CRM-LS-1. The glycan profile was compared with ricin from different preparations and other cultivars and isoforms. A total of 15 different oligomannosidic or paucimannosidic structures were identified in CRM-LS-1. Paucimannose was mainly found within the A-chain and oligomannose constituted the major glycan type of the B-chain. Furthermore, the novel primary structure variants E138 and D138 and four different C-termini of the A-chain as well as two B-chain variants V250 and F250 were elucidated. While the glycan proportions and loci were similar among all variants in CRM-LS-1 and ricin isoforms D and E of all cultivars analysed, a different stoichiometry for isoforms D and E and the amino acid variants were found. This detailed physicochemical characterization of ricin regarding the glycan profile and amino acid sequence variations yields unprecedented insight into the molecular features of this protein toxin. The variable attributes discovered within different cultivars present signature motifs and may allow discrimination of the biotoxin\'s origin that are important in molecular forensic profiling. In conclusion, our data of in-depth CRM-LS-1 characterization combined with the analysis of other cultivars is representative for known ricin variants.
摘要：
EuroBioTox联盟生产了蓖麻毒素（CRM-LS-1）的认证参考材料，以标准化该生物毒素的分析。这项研究建立了蓖麻毒素CRM-LS-1的N-聚糖结构和比例，包括其基因座和占有率。将聚糖谱与来自不同制剂和其他品种和同工型的蓖麻毒素进行了比较。在CRM-LS-1中鉴定出总共15种不同的寡聚体或寡聚体结构。毛甘露糖主要存在于A链内,而寡甘露糖构成B链的主要聚糖类型。此外，阐明了新的一级结构变体E138和D138和A链的四个不同的C末端以及两个B链变体V250和F250。虽然所有品种的CRM-LS-1和蓖麻毒蛋白同工型D和E的所有变体中的聚糖比例和基因座相似，发现了同工型D和E以及氨基酸变体的不同化学计量。蓖麻毒素关于聚糖谱和氨基酸序列变异的这种详细的物理化学表征产生了对这种蛋白质毒素的分子特征的前所未有的了解。在不同品种中发现的可变属性呈现特征基序，并可能允许区分生物毒素的起源，这在分子法医分析中很重要。总之,我们深入的CRM-LS-1表征数据结合对其他品种的分析,对于已知的蓖麻毒素变体具有代表性.
