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首页> 美国卫生研究院文献>International Journal of Molecular Sciences >Predominant Expression of Hybrid N-Glycans Has Distinct Cellular Roles Relative to Complex and Oligomannose N-Glycans
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Predominant Expression of Hybrid N-Glycans Has Distinct Cellular Roles Relative to Complex and Oligomannose N-Glycans

机译：杂种N-聚糖的主要表达具有相对于复杂和寡甘露糖N-聚糖的不同的细胞作用

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Glycosylation modulates growth, maintenance, and stress signaling processes. Consequently, altered N-glycosylation is associated with reduced fitness and disease. Therefore, expanding our understanding of N-glycans in altering biological processes is of utmost interest. Herein, clustered regularly interspaced short palindromic repeats/caspase9 (CRISPR/Cas9) technology was employed to engineer a glycosylation mutant Chinese Hamster Ovary (CHO) cell line, K16, which expresses predominantly hybrid type N-glycans. This newly engineered cell line enabled us to compare N-glycan effects on cellular properties of hybrid type N-glycans, to the well-established Pro−5 and Lec1 cell lines, which express complex and oligomannose types of N-glycans, respectively. Lectin binding studies revealed the predominant N-glycan expressed in K16 is hybrid type. Cell dissociation and migration assays demonstrated the greatest strength of cell–cell adhesion and fastest migratory rates for oligomannose N-glycans, and these properties decreased as oligomannose type were converted to hybrid type, and further decreased upon conversion to complex type. Next, we examined the roles of three general types of N-glycans on ectopic expression of E-cadherin, a cell–cell adhesion protein. Microscopy revealed more functional E-cadherin at the cell–cell border when N-glycans were oligomannose and these levels decreased as the oligomannose N-glycans were processed to hybrid and then to complex. Thus, we provide evidence that all three general types of N-glycans impact plasma membrane architecture and cellular properties.
机译：糖基化调节生长，维持和应激信号传导过程。因此，改变的N-糖基化与降低的适应性和疾病相关。因此，在改变生物过程中扩展对N-聚糖的理解是最重要的。在本文中，采用聚类的规则间隔的短回文重复序列/ caspase9（CRISPR / Cas9）技术来改造糖基化突变体中国仓鼠卵巢（CHO）细胞系K16，该细胞系主要表达杂种N型聚糖。这种新设计的细胞系使我们能够将N-聚糖对杂合型N-聚糖的细胞特性的影响与成熟的Pro - 5和Lec1细胞系进行比较，后者表达复杂的和低聚甘露糖类型。 N-聚糖。凝集素结合研究表明，K16中表达的主要N-聚糖是杂合型。细胞解离和迁移分析表明，低聚甘露糖N-聚糖具有最大的细胞-细胞粘附强度和最快的迁移速率，这些特性随着低聚甘露糖类型转换为杂合型而降低，并在转换为复杂型时进一步降低。接下来，我们研究了三种一般类型的N-聚糖对E-cadherin（一种细胞间粘附蛋白）的异位表达的作用。显微镜检查显示，当N-聚糖为低聚甘露糖时，在细胞-细胞边界处具有更多的功能性E-钙粘着蛋白，随着低聚甘露糖N-聚糖被加工成杂化然后复合的过程，这些水平降低。因此，我们提供了证据，所有三种常规类型的N-聚糖均会影响质膜的结构和细胞特性。

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