N-Glycoproteomic characterization of mannosidase and xylotransferase Mutant Strains of Chlamydomonas.
N-Glycoproteomic characterization of mannosidase and xylotransferase Mutant Strains of Chlamydomonas.
复制标题
衣藻甘露糖苷酶和木糖转移酶突变菌株的 N-糖蛋白组学特征。
DOI：
10.1104/pp.17.01450
复制
发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Hippler Michael
中科院分区：
文献类型：
--
作者：
Schulze Stefan;Oltmanns Anne;Machnik Nick;Liu Gai;Xu Nannan;Jarmatz Niklas;Scholz Martin;Sugimoto Kazuhiko;Fufezan Christian;Huang Kaiyao;Hippler Michael
目前，只有很少的酶机制所需的N-糖基化在衣原体reinhardtii，导致形成的N-聚糖窝藏Xyl和甲基化的Man。reinhardtii N-聚糖不依赖于β 1，2-N-乙酰葡糖胺基转移酶I。在这里，我们进行了比较N-糖蛋白质组学分析的甘露糖苷酶1A（IMMan 1A）和木糖基转移酶1A（IMXylT 1A）的插入突变体。Man 1A的破坏影响Man的甲基化和末端Xyl的添加。与野生型相比，XylT 1A的缺失导致较短的N-聚糖。使用IMMan 1AxIMXylT 1Adouble突变体揭示了Man 1A的缺失抑制了IMXylT 1A表型，表明增加的N-聚糖修剪受核心β 1，2-Xyl调节并且依赖于Man 1A活性。这些数据指向C. reinhardtii与Man 1A和XylT 1A的相互关联的作用。本文所述的结果代表了对C中酶促N-糖基化机制进行功能表征的第一步。莱因哈德氏菌
At present, only little is known about the enzymatic machinery required forN-glycosylation inChlamydomonas reinhardtii, leading to the formation ofN-glycans harboring Xyl and methylated Man. This machinery possesses new enzymatic features, asC. reinhardtii N-glycans are independent of β1,2-N-acetylglucosaminyltransferase I. Here we have performed comparativeN-glycoproteomic analyses of insertional mutants of mannosidase 1A (IMMan1A) and xylosyltransferase 1A (IMXylT1A). The disruption ofman1Aaffected methylation of Man and the addition of terminal Xyl. The absence of XylT1A led to shorterN-glycans compared to the wild type. The use of a IMMan1AxIMXylT1Adouble mutant revealed that the absence of Man1A suppressed the IMXylT1Aphenotype, indicating that the increasedN-glycan trimming is regulated by core β1,2-Xyl and is dependent on Man1A activity. These data point toward an enzymatic cascade in theN-glycosylation pathway ofC. reinhardtiiwith interlinked roles of Man1A and XylT1A. The results described herein represent the first step toward a functional characterization of the enzymaticN-glycosylation machinery inC. reinhardtii.