N-glycosylation is crucial for trafficking and stability of SLC3A2 (CD98).
N-glycosylation is crucial for trafficking and stability of SLC3A2 (CD98).
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DOI：
10.1038/s41598-022-18779-4
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发表时间：
2022-08-26
影响因子：
4.6
通讯作者：
Indiveri, Cesare
中科院分区：
文献类型：
--
作者：
Console, Lara;Scalise, Mariafrancesca;Salerno, Simona;Scanga, Raffaella;Giudice, Deborah;De Bartolo, Loredana;Tonazzi, Annamaria;Indiveri, Cesare
II型糖蛋白CD98(SLC3A2)是一种膜蛋白，在细胞中具有多种作用，包括调节炎症过程、宿主与病原体的相互作用以及与SLC7家族的膜转运蛋白的结合。最近对LAT1复合物中CD98结构的解析表明，四个ASN残基N365、N381、N424、N506含有N-糖基化部分。然后，结合生物信息学、定点突变和细胞生物学方法，研究了N-糖基化对CD98转运和稳定性的影响。这四个ASN的单、双、三和四个突变体的电泳迁移率发生了变化，表观分子质量从95到70 kDa。四重突变体显示了一条70 kDa的单条带，对应于未糖化的蛋白质。分别用生物素化实验和布雷菲尔丁实验检测CD98在细胞膜中的存在和转运。综上所述，结果强调了四重突变严重损害了CD98的稳定性和向质膜的转运。CD98在质膜上的存在减少，与LAT1(SLC7A5)的存在及其转运活性降低有关。这一发现为人类治疗开辟了新的视角。事实上，抑制CD98的运输将与LAT1抑制剂协同作用，这些抑制剂正在进行抗癌治疗的临床试验。
The type II glycoprotein CD98 (SLC3A2) is a membrane protein with pleiotropic roles in cells, ranging from modulation of inflammatory processes, host–pathogen interactions to association with membrane transporters of the SLC7 family. The recent resolution of CD98 structure in complex with LAT1 showed that four Asn residues, N365, N381, N424, N506, harbour N-glycosylation moieties. Then, the role of N-glycosylation on CD98 trafficking and stability was investigated by combining bioinformatics, site-directed mutagenesis and cell biology approach. Single, double, triple and quadruple mutants of the four Asn exhibited altered electrophoretic mobility, with apparent molecular masses from 95 to 70 kDa. The quadruple mutant displayed a single band of 70 kDa corresponding to the unglycosylated protein. The presence in the membrane and the trafficking of CD98 were evaluated by a biotinylation assay and a brefeldin assay, respectively. Taken together, the results highlighted that the quadruple mutation severely impaired both the stability and the trafficking of CD98 to the plasma membrane. The decreased presence of CD98 at the plasma membrane, correlated with a lower presence of LAT1 (SLC7A5) and its transport activity. This finding opens new perspectives for human therapy. Indeed, the inhibition of CD98 trafficking would act synergistically with LAT1 inhibitors that are under clinical trial for anticancer therapy.
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影响因子：
11.2
作者：
Estrach S;Lee SA;Boulter E;Pisano S;Errante A;Tissot FS;Cailleteau L;Pons C;Ginsberg MH;Féral CC
通讯作者：
Féral CC
DOI：
10.1016/j.biocel.2016.11.005
发表时间：
2016-12-01
影响因子：
4
作者：
Ip, Hugh;Sethi, Tariq
通讯作者：
Sethi, Tariq
影响因子：
4.8
作者：
Fort, Joana;de la Ballina, Laura R.;Palacin, Manuel
通讯作者：
Palacin, Manuel
影响因子：
--
作者：
D'Agostino S;Lanzillotta D;Varano M;Botta C;Baldrini A;Bilotta A;Scalise S;Dattilo V;Amato R;Gaudio E;Paduano F;Palmieri C;Iuliano R;Perrotti N;Indiveri C;Fusco A;Gaspari M;Trapasso F
通讯作者：
Trapasso F
DOI：
10.1073/pnas.0404852102
发表时间：
2005-01-11
影响因子：
11.1
作者：
Feral, CC;Nishiya, N;Ginsberg, MH
通讯作者：
Ginsberg, MH