Raft.1, A Monoclonal Antibody Raised Against the Raft Microdomain, Recognizes G-Protein β1 and 2, Which Assemble Near Nucleus After Shiga Toxin Binding to Human Renal Cell Line
Raft.1, A Monoclonal Antibody Raised Against the Raft Microdomain, Recognizes G-Protein β1 and 2, Which Assemble Near Nucleus After Shiga Toxin Binding to Human Renal Cell Line
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Raft.1 是一种针对 Raft 微域的单克隆抗体，可识别志贺毒素与人肾细胞系结合后在细胞核附近组装的 G 蛋白 β1 和 2
DOI：
10.1097/01.lab.0000044134.23951.c8
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发表时间：
2002
影响因子：
5
通讯作者：
J. Fujimoto
中科院分区：
文献类型：
--
作者：
Y. Katagiri;K. Ohmi;Wei;H. Takenouchi;T. Taguchi;N. Kiyokawa;J. Fujimoto
RAFT微区是参与信号转导的富含糖脂的微区支架分子。志贺毒素与人肾小管上皮细胞系ACHN RAFT微区中球状三糖神经酰胺的结合导致了Src-Kinase的瞬时激活。为了研究导致Yes激活的下游信号机制，我们提出了抗RAFT微域的单抗。通过斑点印迹免疫染色结合RAFT微区、渗透后流式细胞术检测细胞内抗原表位、免疫组织化学染色检测STX处理后抗原分子的移位来筛选单抗。Raft.1单抗与STX处理后聚集在细胞核附近特定区域的分子结合。双向免疫印迹和基质辅助激光解吸电离飞行时间质谱仪分析表明，该抗原分子为GTP结合蛋白β亚基1和2(Gβ1和2)。与重组Gβ1和2蛋白的反应性进一步证实了Raft1识别Gβ1和2。据我们所知，这是首次报道产生识别Gβ1和2的单抗。由于Gβ1和2在整个生物体中高度保守，并深入参与信号转导，因此Raft1有望在研究中频繁使用。
Raft microdomains are glycolipid-enriched microdomain scaffolding molecules involved in signal transduction. The binding of Shiga toxin to globotriaosyl ceramide in raft microdomains of the human renal tubular cell line ACHN causes temporal activation of Src-kinase Yes. To study the downstream signaling mechanism proceeding to the activation of Yes, we raised monoclonal antibodies (MAbs) against raft microdomains. The MAbs were screened on the basis of, first, binding to raft microdomains with dot-blot immunostaining, second, intracellular localization of the epitope by flowcytometry after permeabilization, and third, translocation of the antigen molecules after Stx treatment by immunohistochemical staining. Raft.1 MAb bound to the molecules that accumulated to the particular region near the nucleus after Stx treatment. Two-dimensional Western blotting and matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis revealed that the antigen molecule is GTP binding protein β subunits 1 and 2 (Gβ1 and 2). That Raft.1 recognized Gβ1 and 2 was further confirmed by the reactivity to recombinant Gβ1 and 2 proteins. To our knowledge, this is the first report of production of a MAb recognizing Gβ1 and 2. Because Gβ1 and 2 are highly conserved all through organisms and are deeply involved in signal transduction, Raft.1 is expected to be utilized frequently in research.
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影响因子：
4.8
作者：
Y. Toya;C. Schwencke;J. Couet;M. Lisanti;Y. Ishikawa
通讯作者：
Y. Toya;C. Schwencke;J. Couet;M. Lisanti;Y. Ishikawa
影响因子：
2.9
作者：
Pike, LJ;Han, XL;Gross, RW
通讯作者：
Gross, RW
牛视网膜杆外节转导蛋白的表征。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Fung,BK
通讯作者：
Fung,BK
哺乳动物 G 蛋白 β 亚基家族的第五个成员。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Watson,AJ;Katz,A;Simon,MI
通讯作者：
Simon,MI
影响因子：
2.9
作者：
Ahmed, SN;Brown, DA;London, E
通讯作者：
London, E