Lysine at position 329 within a C-terminal dilysine motif is crucial for the ER localization of human SLC35B4.

Lysine at position 329 within a C-terminal dilysine motif is crucial for the ER localization of human SLC35B4.
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DOI:
10.1371/journal.pone.0207521
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发表时间:
2018
期刊:
影响因子:
3.7
通讯作者:
Olczak M
Olczak M
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Bazan B;Wiktor M;Maszczak-Seneczko D;Olczak T;Kaczmarek B;Olczak M

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SLC 35 B4属于溶质载体35(SLC 35)家族,其最佳表征的成员显示核苷酸糖转运活性。利用HepG 2细胞实验模型和间接免疫荧光染色,我们验证了SLC 35 B4定位于内质网(ER)。我们证明,二赖氨酸基序,特别是赖氨酸在位置329,是至关重要的ER定位的这种蛋白质在人类细胞中,因此,应该谨慎使用蛋白质C-标记。为了验证蛋白质在糖缀合物合成中的重要性,我们使用CRISPR-Cas9方法产生了SLC 35 B4缺陷型HepG 2细胞系。我们的数据显示,SLC 35 B4基因的敲除不影响主要的UDP-Xyl-和UDP-GlcNAc-依赖性糖基化途径。
SLC35B4 belongs to the solute carrier 35 (SLC35) family whose best-characterized members display a nucleotide sugar transporting activity. Using an experimental model of HepG2 cells and indirect immunofluorescent staining, we verified that SLC35B4 was localized to the endoplasmic reticulum (ER). We demonstrated that dilysine motif, especially lysine at position 329, is crucial for the ER localization of this protein in human cells and therefore one should use protein C-tagging with caution. To verify the importance of the protein in glycoconjugates synthesis, we generated SLC35B4-deficient HepG2 cell line using CRISPR-Cas9 approach. Our data showed that knock-out of the SLC35B4 gene does not affect major UDP-Xyl- and UDP-GlcNAc-dependent glycosylation pathways.
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