Sora I, Richman J, Santoro G, Wei H, Wang Y, Vanderah T, Horvath R, Nguyen M, Waite S, Roeske W R
Department of Pharmacology, College of Medicine, University of Arizona, Tucson 85724.
Biochem Biophys Res Commun. 1994 Oct 14;204(1):419-27. doi: 10.1006/bbrc.1994.2475.
A human creatine transporter (hCRT-BS2M) cDNA clone was isolated from a human brainstem/spinal cord using a PCR and phage plaque hybridization based technique. This clone included an open reading frame of 1,905 base pairs(bp) within a 2,283bp cDNA. Northern blot hybridization detected the expression of corresponding mRNAs most prominently in the skeletal muscle, heart and kidney. Peptide sequence analysis of the hCRT-BS2M protein product revealed 12 putative transmembrane domains. The predicted protein sequence further demonstrates that the hCRT-BS2M has highly conserved amino acid identity with the other members of the sodium dependent plasma membrane transporter family. Transient expression of the hCRT-BS2M in COS-7 cells demonstrates sodium dependent [14C]creatine uptake with a KM value of 14.9 +/- 3.0 microM (n = 5) that is attenuated by creatine and selective structural analogues of creatine.
采用基于聚合酶链反应（PCR）和噬菌体菌斑杂交的技术，从人脑干/脊髓中分离出一个人类肌酸转运体（hCRT - BS2M）cDNA克隆。该克隆在一个2283bp的cDNA内包含一个1905个碱基对（bp）的开放阅读框。Northern印迹杂交检测到相应的mRNA在骨骼肌、心脏和肾脏中表达最为显著。对hCRT - BS2M蛋白产物的肽序列分析揭示了12个推定的跨膜结构域。预测的蛋白质序列进一步表明，hCRT - BS2M与钠依赖性质膜转运体家族的其他成员具有高度保守的氨基酸同一性。hCRT - BS2M在COS - 7细胞中的瞬时表达显示出钠依赖性的[14C]肌酸摄取，其KM值为14.9±3.0微摩尔（n = 5），肌酸和肌酸的选择性结构类似物可使其摄取减弱。
