Expression of the rat brain creatine transporter in situ and in transfected HeLa cells.
Expression of the rat brain creatine transporter in situ and in transfected HeLa cells.
复制标题
大鼠脑肌酸转运蛋白在原位和转染的 HeLa 细胞中的表达。
DOI：
10.1159/000111450
复制
发表时间：
1996
影响因子：
2.9
通讯作者：
Blakely,RD
中科院分区：
文献类型：
--
作者：
Saltarelli,MD;Bauman,AL;Moore,KR;Bradley,CC;Blakely,RD
利用编码γ-氨基丁酸/去甲肾上腺素转运蛋白基因家族保守区的简并寡核苷酸探针，克隆了大鼠脑肌酸转运蛋白基因。RCREAT编码一个高度疏水的635个氨基酸的蛋白质，具有12个潜在的跨膜结构域和典型的N-连接糖基化和蛋白质磷酸化位点。将rCREAT基因导入哺乳动物细胞，可导致[14C]肌酸摄取的表达，此作用可被低摩尔浓度的肌酸摄取抑制剂所阻断。两个rCREAT mRNAs在大鼠的大脑、视网膜、肾脏和心脏中表达。全脑rCREAT mRNAs显示，出生后显著上升到稳定的成人水平。原位杂交研究表明，rCREAT在成年大鼠脑内广泛而不同地表达，在有髓纤维束、小脑颗粒细胞、海马锥体细胞、脑干核团和脉络丛内皮细胞中高表达。这些研究将有助于开发新的分子探针，用于确定肌酸转运蛋白的细胞表达模式、在ATP动态平衡中的功能以及与细胞能量代谢障碍的关系。
Using degenerate oligonucleotide probes encoding conserved regions of the γ-aminobutyric acid/norepinephrine transporter gene family, we have cloned a rat brain cDNA encoding a creatine transporter (rCREAT). rCREAT encodes a highly hydrophobic, 635-amino-acid protein possessing 12 potential transmembrane domains and canonical sites for N-linked glycosylation and protein phosphorylation. Transfection of rCREAT cDNA into mammalian cells results in the expression of [14C]creatine uptake, which is blocked by low micromolar concentrations of recognized creatine uptake inhibitors. Two rCREAT mRNAs are expressed in the rat brain, retina, kidney and heart. Whole-brain rCREAT mRNAs demonstrate a marked postnatal rise to steady-state adult levels. In situ hybridization studies indicate a widespread, differential rCREAT mRNA expression in adult rat brain, with high expression noted over myelinated fiber tracts, cerebellar granule cells, hippocampal pyramidal cells, brainstem nuclei and endothelial cells of the choroid plexus. These studies will allow the development of new molecular probes useful for defining the creatine transporter's cellular expression pattern, function in ATP homeostasis and association with disorders of cellular energy metabolism.