Altered N-glycan composition impacts flagella-mediated adhesion in Chlamydomonas reinhardtii.
Altered N-glycan composition impacts flagella-mediated adhesion in Chlamydomonas reinhardtii.
复制标题
改变的N-聚糖组成影响莱茵衣藻鞭毛介导的粘附。
DOI：
10.7554/elife.58805
复制
发表时间：
2020-12-10
期刊：
影响因子：
7.7
通讯作者：
Hippler M
中科院分区：
文献类型：
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作者：
Xu N;Oltmanns A;Zhao L;Girot A;Karimi M;Hoepfner L;Kelterborn S;Scholz M;Beißel J;Hegemann P;Bäumchen O;Liu LN;Huang K;Hippler M
对于单细胞的莱茵衣藻，N-糖基化蛋白的存在下，两个鞭毛的表面是至关重要的细胞间的相互作用，在交配和鞭毛表面粘附。然而，尚不清楚是否只有与相应蛋白质连接的N-聚糖的存在或组成对于这些过程是重要的。为此，我们测试了几个C。在一些实施方案中，所述突变体包括莱茵衣藻插入突变体和木糖基转移酶IA的CRISPR/Cas9敲除突变体，所有这些突变体都具有改变的N-聚糖组成。利用原子力显微镜和微量移液器力测量，我们的数据显示，减少N-聚糖的复杂性阻碍了所需的粘附力结合鞭毛的表面。这导致聚苯乙烯珠结合和转运受损，但细胞在固体表面上的滑动不受影响。值得注意的是，组装，鞭毛内运输，和蛋白质输入到鞭毛不受改变的N-糖基化。因此，我们得出结论，适当的N-糖基化的鞭毛蛋白是至关重要的粘附C。reinhardtii细胞粘附到表面上，表明N-聚糖通过直接表面接触介导表面粘附。
For the unicellular alga Chlamydomonas reinhardtii, the presence of N-glycosylated proteins on the surface of two flagella is crucial for both cell-cell interaction during mating and flagellar surface adhesion. However, it is not known whether only the presence or also the composition of N-glycans attached to respective proteins is important for these processes. To this end, we tested several C. reinhardtii insertional mutants and a CRISPR/Cas9 knockout mutant of xylosyltransferase 1A, all possessing altered N-glycan compositions. Taking advantage of atomic force microscopy and micropipette force measurements, our data revealed that reduction in N-glycan complexity impedes the adhesion force required for binding the flagella to surfaces. This results in impaired polystyrene bead binding and transport but not gliding of cells on solid surfaces. Notably, assembly, intraflagellar transport, and protein import into flagella are not affected by altered N-glycosylation. Thus, we conclude that proper N-glycosylation of flagellar proteins is crucial for adhering C. reinhardtii cells onto surfaces, indicating that N-glycans mediate surface adhesion via direct surface contact.