Marie Hélène, Billups Daniela, Bedford Fiona K, Dumoulin Andrea, Goyal Rakesh K, Longmore Gregory D, Moss Stephen J, Attwell David
Department of Physiology, Laboratory of Molecular Cell Biology, University College London, Gower Street, London, WC1E 6BT, United Kingdom.
Mol Cell Neurosci. 2002 Feb;19(2):152-64. doi: 10.1006/mcne.2001.1066.
We have identified a cytoplasmic LIM protein, Ajuba, which interacts with the amino terminus of GLT-1, the most abundant plasma membrane glutamate transporter in the brain. Ajuba has a cytoplasmic location when expressed alone in COS cells, but translocates to colocalize with GLT-1 at the plasma membrane when GLT-1 is coexpressed. Ajuba is expressed in cerebellum, cortex, hippocampus, and retina and also in organs outside the CNS. Ajuba is found with GLT-1 in astrocytes, cerebellar Bergmann glia and retinal neurons, and antibodies to Ajuba coimmunoprecipitate GLT-1 from brain. For GLT-1 expressed in COS cells, coexpression of Ajuba did not affect the transporter's K(m) or V(max) for glutamate. Since Ajuba is known to activate MAP kinase enzymes, and its homologue Zyxin binds to cytoskeletal proteins, we propose that Ajuba is a scaffolding protein allowing GLT-1 to regulate intracellular signaling or interact with the cytoskeleton.
我们鉴定出一种细胞质LIM蛋白Ajuba,它与GLT-1的氨基末端相互作用,GLT-1是大脑中最丰富的质膜谷氨酸转运体。单独在COS细胞中表达时,Ajuba位于细胞质中,但当与GLT-1共表达时,它会转位至质膜并与GLT-1共定位。Ajuba在小脑、皮质、海马体和视网膜以及中枢神经系统以外的器官中均有表达。在星形胶质细胞、小脑伯格曼胶质细胞和视网膜神经元中发现Ajuba与GLT-1共存,并且针对Ajuba的抗体可从大脑中共免疫沉淀GLT-1。对于在COS细胞中表达的GLT-1,Ajuba的共表达不会影响转运体对谷氨酸的K(m)或V(max)。由于已知Ajuba可激活丝裂原活化蛋白激酶(MAP激酶),且其同源物斑联蛋白可与细胞骨架蛋白结合,因此我们推测Ajuba是一种支架蛋白,可使GLT-1调节细胞内信号传导或与细胞骨架相互作用。