Ganapathy V, Ramamoorthy J D, Del Monte M A, Leibach F H, Ramamoorthy S
Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta 30912-2100.
Curr Eye Res. 1995 Sep;14(9):843-50. doi: 10.3109/02713689508995807.
This investigation was undertaken to study the role of cAMP in the regulation of the taurine transporter expressed in a human retinal pigment epithelial (HRPE) cell line. Treatment of the HRPE cells with cholera toxin for 24 h was found to stimulate the taurine transporter activity, as measured by taurine transport into the cells in the presence of NaCl, to a significant extent. The stimulation was 50-60% at 100 ng/ml cholera toxin. This stimulation was specific to the taurine transporter since the transport of two other amino acids (leucine and alanine), which are not substrates for the taurine transporter, was not affected by cholera toxin under similar conditions. Exposure of the cells to cholera toxin for a time period > 4 h was needed to elicit the stimulatory effect. The cholera toxin-induced stimulation of the taurine transporter activity was associated with an increase in the maximal velocity of the transport system. The affinity of the transporter for taurine was not altered by the treatment. The stimulatory effect was markedly blunted when the treatment of the cells with cholera toxin was done in the presence of actinomycin D, an inhibitor of transcription, or cycloheximide, an inhibitor of translation. The increase in the taurine transporter activity induced by cholera toxin was associated with a 2.6-fold increase in the steady state levels of the transporter mRNA. Measurement of cyclic nucleotides in control and cholera toxin-treated cells revealed that the toxin caused a 20-fold increase in the cellular levels of cAMP, the levels of cGMP remaining unaffected.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究旨在探讨环磷酸腺苷(cAMP)在调控人视网膜色素上皮(HRPE)细胞系中表达的牛磺酸转运体方面的作用。结果发现,用霍乱毒素处理HRPE细胞24小时可显著刺激牛磺酸转运体活性,这一活性通过在氯化钠存在的情况下牛磺酸进入细胞的转运来测定。在100 ng/ml霍乱毒素作用下,刺激率为50 - 60%。这种刺激对牛磺酸转运体具有特异性,因为在类似条件下,另外两种不是牛磺酸转运体底物的氨基酸(亮氨酸和丙氨酸)的转运不受霍乱毒素影响。细胞暴露于霍乱毒素超过4小时才会引发刺激作用。霍乱毒素诱导的牛磺酸转运体活性刺激与转运系统最大速度的增加有关。处理并未改变转运体对牛磺酸的亲和力。当在转录抑制剂放线菌素D或翻译抑制剂环己酰亚胺存在的情况下用霍乱毒素处理细胞时,刺激作用明显减弱。霍乱毒素诱导的牛磺酸转运体活性增加与转运体mRNA稳态水平增加2.6倍有关。对照细胞和经霍乱毒素处理的细胞中环核苷酸的测量结果显示,毒素使细胞内cAMP水平增加了20倍,而cGMP水平未受影响。(摘要截短至250字)