Ito K, Groudine M
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104, USA.
J Biol Chem. 1997 Oct 17;272(42):26780-6. doi: 10.1074/jbc.272.42.26780.
We have isolated and characterized a novel member (CAT3) of the cationic amino acid transporter (CAT) family. In oocyte injection assays, CAT3 cRNA exhibited a saturable, sodium ion-independent transport activity with high affinity for L-arginine and L-lysine (Km = 40-60 and 115-165 microM, respectively). Transport of L-arginine was effectively competed only by cationic amino acids in L-form: arginine, lysine, ornithine, and 2,4-diamino-n-butyric acid but not by 2,3-diaminopropionic acid. The presence of L-arginine in the incubation medium stimulated the efflux rate of L-arginine, indicating that CAT3 is subject to trans-stimulation. All these results are consistent with the idea that CAT3, along with CAT1 and CAT2, constitutes the transport activity originally assigned to system y+. Like CAT2, but unlike CAT1, the expression of CAT3 is regulated in a highly tissue-specific manner; when various adult tissues were examined, significant levels of CAT3 transcript were detectable only in brain. In situ hybridization on brain sections revealed that CAT3 transcripts were localized predominantly along the midbrain-thalamus-hypothalamus axis, whereas neither CAT1 nor CAT2 transcripts demonstrated a similar localization. In contrast to its highly localized expression during the primitive streak stage and in the adult stage, CAT3 expression was detected more widely in 13.5 day post-coitum mouse embryos.
我们分离并鉴定了阳离子氨基酸转运体(CAT)家族的一个新成员(CAT3)。在卵母细胞注射试验中,CAT3的cRNA表现出一种可饱和的、不依赖钠离子的转运活性,对L-精氨酸和L-赖氨酸具有高亲和力(Km分别为40 - 60和115 - 165 microM)。L-精氨酸的转运仅被L型阳离子氨基酸有效竞争:精氨酸、赖氨酸、鸟氨酸和2,4-二氨基正丁酸,但不被2,3-二氨基丙酸竞争。孵育培养基中L-精氨酸的存在刺激了L-精氨酸的流出速率,表明CAT3受到反式刺激。所有这些结果都与以下观点一致,即CAT3与CAT1和CAT2一起构成了最初分配给系统y +的转运活性。与CAT2一样,但与CAT1不同,CAT3的表达以高度组织特异性的方式受到调节;当检查各种成年组织时,仅在脑中可检测到显著水平的CAT3转录本。脑切片的原位杂交显示,CAT3转录本主要定位于中脑 - 丘脑 - 下丘脑轴,而CAT1和CAT2转录本均未显示出类似的定位。与其在原条期和成年期高度局限的表达相反,在妊娠13.5天的小鼠胚胎中检测到更广泛的CAT3表达。