David V, Hochstenbach F, Rajagopalan S, Brenner M B
Department of Rheumatology and Immunology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115.
J Biol Chem. 1993 May 5;268(13):9585-92.
A cDNA clone encoding the human endoplasmic reticulum (ER) resident protein IP90 was isolated and sequenced. It predicts a transmembrane protein with a large ER luminal region showing sequence similarity to calreticulin and a short cytoplasmic domain containing a COOH-terminal RKPRRE sequence that may be relevant to its retention in the ER. It is 95% homologous to the canine ER membrane phosphorprotein called pp90 or calnexin (Wada, I., Rindress, D., Cameron, P. H., Ou, W.-J., Doherty, J. J., II, Louvard, D., Bell, A. W., Dignard, D., Thomas, D. Y., and Bergeron, J. J. M. (1991) J. Biol. Chem. 266, 19599-19610). Previously, in lymphocytes, we have characterized IP90 as a protein associated with partially assembled multichain proteins including the T cell receptor, the membrane immunoglobulin, and the heavy chain of the major histocompatibility complex class I molecules (Hochstenbach, F., David, V., Watkins, S., and Brenner, M. B. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 4734-4738). Here, we show that within a short metabolic labeling period, IP90 associates transiently with many different newly synthesized proteins. However, in a T cell line that cannot assemble a complete T cell receptor because it lacks the alpha subunit, the unassembled T cell receptor beta chains, which are retained in the ER, remain associated with IP90 throughout a prolonged chase time period. Together, these data offer further evidence suggesting that IP90 may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits.
分离并测序了一个编码人内质网(ER)驻留蛋白IP90的cDNA克隆。它预测的跨膜蛋白有一个大的ER腔区域,与钙网蛋白有序列相似性,还有一个短的细胞质结构域,包含一个COOH末端的RKPRRE序列,这可能与其在内质网中的滞留有关。它与犬类内质网膜磷蛋白pp90或钙连蛋白有95%的同源性(和田,I.,林德雷斯,D.,卡梅伦,P. H.,欧,W.-J.,多尔蒂,J. J.,II,卢瓦尔,D.,贝尔,A. W.,迪尼亚德,D.,托马斯,D. Y.,和伯杰龙,J. J. M.(1991)《生物化学杂志》266,19599 - 19610)。此前,在淋巴细胞中,我们已将IP90鉴定为一种与部分组装的多链蛋白相关的蛋白,这些多链蛋白包括T细胞受体、膜免疫球蛋白和主要组织相容性复合体I类分子的重链(霍赫施滕巴赫,F.,大卫,V.,沃特金斯,S.,和布伦纳,M. B.(1992)《美国国家科学院院刊》89,4734 - 4738)。在此,我们表明在短时间的代谢标记期内,IP90与许多不同的新合成蛋白短暂结合。然而,在一个由于缺乏α亚基而无法组装完整T细胞受体的T细胞系中,未组装的、滞留在ER中的T细胞受体β链在延长的追踪时间段内仍与IP90结合。这些数据共同提供了进一步的证据,表明IP90可能在协助蛋白质组装和/或在ER中滞留未组装的蛋白质亚基方面发挥作用。