Differential expression of an E-selectin ligand (SLex) by two Chinese hamster ovary cell lines transfected with the same alpha (1,3)-fucosyltransferase gene (ELFT).
Differential expression of an E-selectin ligand (SLex) by two Chinese hamster ovary cell lines transfected with the same alpha (1,3)-fucosyltransferase gene (ELFT).
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转染相同 α (1,3)-岩藻糖基转移酶基因 (ELFT) 的两种中国仓鼠卵巢细胞系 E-选择素配体 (SLex) 的差异表达。
DOI：
10.1016/s0021-9258(17)42216-2
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
P. Stanley
中科院分区：
文献类型：
--
作者：
S. Goelz;R. Kumar;B. Potvin;S. Sundaram;M. Brickelmaier;P. Stanley
编码α(1，3)-岩藻糖基转移酶(α(1，3)-Fuc-T)的哺乳动物基因被称为Elam-1配体岩藻糖基转移酶(ELFT)或Fuc-TIV。(Goelz，S.E.，Hession，C.，Goff，D.，Griffiths，B.，Tizard，R.，Newman，B.，chi-Rosso，G.和Lobb，R.(1990)Cell 63,1349-1356)发现表达ELFT基因的中国仓鼠卵巢(CHO)细胞具有α(1，3)Fuc-T活性并能与E-选择素结合。相比之下，Lowe等人。(Lowe，J.B.，Kukowska-Latallo，J.F.，Nair，R.P.，Larsen，R.D.，Marks，R.M.，Macher，B.A.，Kelly，R.J.和Ernst，L.K.(1991)J.Biol.化学。17467-17477)和库马尔等人。(Kumar，R.，Potvin，B.，Muller，W.A.和Stanley，P.(1991)J.Biol.化学。21777-21783)发现，表达相同的α(1，3)Fuc-T基因的CHO转基因细胞没有与E-选择素结合；后者也没有合成已知的E-选择素配体唾液酸化Lex(Slex)，尽管它们具有相当大的α(1，3)Fuc-T活性。我们现在表明，这些不同的结果是由于亲本CHO细胞系之间的差异。将ELFT基因导入Pro-5或二氢叶酸还原酶(DHFR)-CHO细胞后，只有DHFR-CHO细胞表达SLeX并与E-选择素结合。来自单抗和凝集素结合研究的间接证据表明，Pro-5和DHFR-CHO细胞系合成的碳水化合物结构范围不同。由于DHFR-/ELFT转基因细胞表达细胞表面的SLeX，但在体外很难将岩藻糖转移到唾液酸化的底物上，因此ELFT可能能够将Pro-5细胞中缺失的复杂碳水化合物岩藻糖基化。或者，任何一种CHO系都可能具有修饰α(1，3)岩藻糖化乳糖胺的活性(如α(2，3)唾液酸基转移酶)。
The mammalian cDNA encoding alpha (1,3)-fucosyltransferase (alpha (1,3)Fuc-T) termed ELAM-1 ligand fucosyltransferase (ELFT) or Fuc-TIV was previously cloned by three groups who reported different results from transfection studies Goelz et al. (Goelz, S. E., Hession, C., Goff, D., Griffiths, B., Tizard, R., Newman, B., Chi-Rosso, G., and Lobb, R. (1990) Cell 63, 1349-1356) found that Chinese hamster ovary (CHO) cells expressing the ELFT cDNA had alpha (1,3)Fuc-T activity and were able to bind to E-selectin. In contrast, Lowe et al. (Lowe, J. B., Kukowska-Latallo, J. F., Nair, R. P., Larsen, R. D., Marks, R. M., Macher, B. A., Kelly, R. J., and Ernst, L. K. (1991) J. Biol. Chem. 266, 17467-17477) and Kumar et al. (Kumar, R., Potvin, B., Muller, W. A., and Stanley, P. (1991) J. Biol. Chem. 266, 21777-21783) found no binding to E-selectin of CHO transfectants expressing the same alpha (1,3)Fuc-T gene; nor did the latter transfectants synthesize a known E-selectin ligand, sialylated Lex (SLex), although they had substantial alpha (1,3)Fuc-T activity. We now show that these discrepant results were due to a difference between the parental CHO cell lines. Following transfection of ELFT cDNA into Pro-5 or dihydrofolate reductase (DHFR)- CHO cells, only the DHFR- transfectants expressed SLex and bound to E-selectin. Indirect evidence from monoclonal antibody and lectin binding studies indicates that the range of carbohydrate structures synthesized by the Pro-5 and DHFR- CHO cell lines differs. Since DHFR-/ELFT transfectants expressed cell surface SLex but transferred fucose poorly to sialylated substrates in vitro, ELFT may be able to fucosylate a complex carbohydrate missing from Pro-5 cells. Alternatively, either CHO line may have an activity (such as an alpha (2,3)-sialyltransferase), that modifies alpha (1,3)-fucosylated lactosamines.