Reconstitution of functional L-selectin ligands on a cultured human endothelial cell line by cotransfection of alpha1-->3 fucosyltransferase VII and newly cloned GlcNAcbeta:6-sulfotransferase cDNA.
Reconstitution of functional L-selectin ligands on a cultured human endothelial cell line by cotransfection of alpha1-->3 fucosyltransferase VII and newly cloned GlcNAcbeta:6-sulfotransferase cDNA.
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通过共转染 α1-->3 岩藻糖基转移酶 VII 和新克隆的 GlcNAcbeta:6-磺基转移酶 cDNA，在培养的人内皮细胞系上重建功能性 L-选择素配体。
DOI：
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发表时间：
1999
影响因子：
11.1
通讯作者：
R. Kannagi
中科院分区：
文献类型：
--
作者：
N. Kimura;C. Mitsuoka;A. Kanamori;N. Hiraiwa;K. Uchimura;T. Muramatsu;T. Tamatani;G. Kansas;R. Kannagi
最近，我们提出唾液酸 6-磺基 Lewis X 作为人类淋巴结高内皮微静脉上 L-选择素配体的主要碳水化合物加帽基团。在这项研究中，我们通过转染 alpha1-->3 岩藻糖基转移酶 VII (Fuc-T VII) 和新克隆的 GlcNAcbeta:6-磺基转移酶 (6-Sul-T) cDNA，成功地在培养的人内皮细胞系 ECV304 上重建功能性 L-选择素配体。转染Fuc-T VII cDNA的ECV304细胞表达传统的唾液酸Lewis X，用包括2H5在内的特异性抗体检测到，而转染6-Sul-T cDNA的细胞表达唾液酸6-磺基乳糖胺以及MECA-79定义的碳水化合物决定簇，但这些单转染的细胞未能表达唾液酸6-磺基Lewis X，如用抗唾液酸6-磺基检测到的路易斯 X 单克隆抗体 G152。唾液酸6-磺基Lewis X仅出现在用6-Sul-T和Fuc-T VII cDNA共转染的细胞上。仅在双转染的 ECV304 细胞中观察到 L-选择素表达细胞的显着粘附，并且被 G152 抑制。未观察到单独用6-Sul-T或Fuc-T VII cDNA转染的细胞的粘附。在人内皮细胞中，两种酶的 mRNA 在白细胞介素 1 刺激下表达或诱导。这些结果表明，通过两种酶的协同作用合成的一组碳水化合物决定簇（通常以唾液酸6-磺基路易斯X封端基团为代表）充当L-选择素配体的重要组成部分，并且在早期研究中用于检测高内皮小静脉上的L-选择素配体的试剂2H5和MECA-79识别同一组合成产物的两个不同方面。
Recently, we proposed sialyl 6-sulfo Lewis X as a major carbohydrate-capping group of the L-selectin ligands on high endothelial venules in human lymph nodes. In this study we succeeded in reconstituting functional L-selectin ligands on a cultured human endothelial cell line, ECV304, by transfecting the alpha1-->3fucosyltranseferase VII (Fuc-T VII) and newly cloned GlcNAcbeta:6-sulfotransferase (6-Sul-T) cDNAs. The ECV304 cells transfected with Fuc-T VII cDNA expressed conventional sialyl Lewis X detected with specific antibodies including 2H5, whereas the cells transfected with 6-Sul-T cDNA expressed sialyl 6-sulfo lactosamine as well as MECA-79-defined carbohydrate determinants, but these singly transfected cells failed to express sialyl 6-sulfo Lewis X, as detected with the antisialyl 6-sulfo Lewis X mAb G152. Sialyl 6-sulfo Lewis X appeared only on the cells that were cotransfected with both 6-Sul-T and Fuc-T VII cDNAs. Significant adhesion of L-selectin-expressing cells was seen only to the doubly transfected ECV304 cells and was inhibited by G152. No adhesion was observed to the cells transfected either with 6-Sul-T or with Fuc-T VII cDNA alone. The mRNAs of the two enzymes were expressed or were inducible upon interleukin 1 stimulation in human endothelial cells. These results indicate that a set of carbohydrate determinants synthesized by the concerted action of the two enzymes, as typically represented by the sialyl 6-sulfo Lewis X-capping group, serves as an essential component of the ligand for L-selectin and that the reagents 2H5 and MECA-79, utilized in earlier studies to detect L-selectin ligand on high endothelial venules, recognize two different aspects of the same set of synthetic products.
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DOI：
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发表时间：
1994-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
S. Natsuka;K. M. Gersten;K. Zenita;R. Kannagi;J. Lowe
通讯作者：
S. Natsuka;K. M. Gersten;K. Zenita;R. Kannagi;J. Lowe
DOI：
10.1073/pnas.87.6.2244
发表时间：
1990-03-01
影响因子：
11.1
作者：
KISHIMOTO, TK;JUTILA, MA;BUTCHER, EC
通讯作者：
BUTCHER, EC
L-选择素-碳水化合物相互作用：路易斯 x 三糖的相关修饰。
影响因子：
2.9
作者：
Sanders,WJ;Katsumoto,TR;Bertozzi,CR;Rosen,SD;Kiessling,LL
通讯作者：
Kiessling,LL
影响因子：
11.2
作者：
K. Fukushima;M. Hirota;P. Terasaki;A. Wakisaka;H. Togashi;D. Chia;N. Suyama;Y. Fukushi;E. Nudelman;S. Hakomori
通讯作者：
K. Fukushima;M. Hirota;P. Terasaki;A. Wakisaka;H. Togashi;D. Chia;N. Suyama;Y. Fukushi;E. Nudelman;S. Hakomori
L-选择素的高内皮小静脉配体：鉴定和功能。
影响因子：
3.9
作者：
Rosen,SD;Hwang,ST;Giblin,PA;Singer,MS
通讯作者：
Singer,MS