Association of Src Family Tyrosine Kinase Lyn with Ganglioside GD3 in Rat Brain
Association of Src Family Tyrosine Kinase Lyn with Ganglioside GD3 in Rat Brain
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大鼠脑中 Src 家族酪氨酸激酶 Lyn 与神经节苷脂 GD3 的关联
DOI：
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发表时间：
1997
影响因子：
4.8
通讯作者：
Y. Sanai
中科院分区：
文献类型：
--
作者：
K. Kasahara;Yumiko Watanabe;Tadashi Yamamoto;Y. Sanai
通过与抗绞痛抗体共免疫沉淀检查中枢神经系统中的神经节剂与特异性蛋白的关联。在大鼠脑的膜分数中，在具有与神经节苷脂GD3（R24）的单克隆抗体的沉淀物中检测到蛋白激酶活性。使用体外激酶测定法，通过凝胶电泳分离出几种40、53、56和80 kDa的磷酸化蛋白。在这些蛋白质中，基于凝胶电泳期间的共迁移，比较肽映射和对抗Lyn抗体的顺序免疫致敬。使用中国仓鼠卵巢（CHO）细胞中的cDNA表达系统证实了该鉴定，该细胞仅表达GM3（GM3），即GD3Synthase的酶促底物。在与GD3合酶和Lyn表达质粒共转染中，R24免疫沉淀的LYN和抗LYN抗体免疫沉淀GD3。 R24大鼠原发小脑培养物的治疗诱导了几种包括有丝分裂原激活蛋白激酶在内的几种底物的LYN激活和快速的酪氨酸磷酸化。此外，蔗糖密度梯度分析表明，小脑和CHO转染剂的lyn在低密度光散射带中被检测到，即口腔膜膜分数。 R24来自CHO转染剂Triton X-100提取物的R24免疫沉淀小窝蛋白。这些观察结果表明，GD3可能在脑细胞膜上的小窝样结构域中调节Lyn。
Association of gangliosides with specific proteins in the central nervous system was examined by co-immunoprecipitation with anti-ganglioside antibody. Protein kinase activity was detected in precipitates with monoclonal antibody to ganglioside GD3 (R24) from membranal fraction of rat brain. Using in vitro kinase assay, several phosphorylated proteins of 40, 53, 56, and 80 kDa were isolated by gel electrophoresis. Of these proteins, the proteins of 53 and 56 kDa (p53/56) were identified as two isoforms of Src family tyrosine kinase Lyn, based on co-migration during gel electrophoresis, comparative peptide mapping, and sequential immunoprecipitation with anti-Lyn antibody. The identification was confirmed using a cDNA expression system in Chinese hamster ovary (CHO) cells, which express solely ganglioside GM3, the enzymatic substrate of GD3synthase. In co-transfection with GD3 synthase and Lyn expression plasmids, R24 immunoprecipitated Lyn and anti-Lyn antibody immunoprecipitated GD3. R24 treatment of rat primary cerebellar cultures induced Lyn activation and rapid tyrosine phosphorylation of several substrates including mitogen-activated protein kinases. Furthermore, sucrose density gradient analysis showed that Lyn of cerebellum and CHO transfectants were detected in a low density light-scattering band, i.e. the caveolae membrane fraction. R24 immunoprecipitated caveolin from Triton X-100 extract of CHO transfectants. These observations suggest that GD3 may regulate Lyn in a caveolae-like domain on brain cell membranes.
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影响因子：
56.9
作者：
GRANT, SGN;ODELL, TJ;KANDEL, ER
通讯作者：
KANDEL, ER
DOI：
10.1073/pnas.91.25.12130
发表时间：
1994-12-06
影响因子：
11.1
作者：
SCHROEDER, R;LONDON, E;BROWN, D
通讯作者：
BROWN, D
抗 GD3 神经节苷脂单克隆抗体刺激 T 淋巴细胞增殖。
DOI：
--
发表时间：
1987
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Welte,K;Miller,G;Chapman,PB;Yuasa,H;Natoli,E;Kunicka,JE;Cordon-Cardo,C;Buhrer,C;Old,LJ;Houghton,AN
通讯作者：
Houghton,AN
影响因子：
4.8
作者：
Li, SW;Couet, J;Lisanti, MP
通讯作者：
Lisanti, MP
每个小凹包含多个糖基磷脂酰肌醇锚定膜蛋白。
DOI：
10.1101/sqb.1992.057.01.065
发表时间：
1992
期刊：
Cold Spring Harbor symposia on quantitative biology
影响因子：
--
作者：
Ying,YS;Anderson,RG;Rothberg,KG
通讯作者：
Rothberg,KG