Structural and functional analysis of a cystine/glutamate-specific carrier protein of plasma membrane

质膜胱氨酸/谷氨酸特异性载体蛋白的结构和功能分析

• 批准号: 04670136
• 负责人: ISHII Tetsuro
• 金额: $ 1.34万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670136/
• 关键词: Cystine; Stress protein; Amino acid transport; cDNA; Macrophage; グルタミン酸

Stress agents induces cystine transport activity in mouse macrophages cultured in vitro. The induction of cystine entry into the cells enhances synthesis of glutathione, an anti-oxidative cellular substance. We tried to clone the carrier of cystine by the differential screening to select stress-induced clones. We constructed a directional cDNA library from 1.5-3 kb fraction of mouse macrophage mRNA.We selected total 800 clones from about 8x10^4 phage plaques. We could not detect any expression of the cystine transport activity in Xenopus laevis oocytes after microinjection of cRNAs synthesized from those cDNAs.Although we could not isolate a cNDA clone encodnig cystine carrier, we have succeeded in the cloning of a stress-induced 23 kDa protein named MSP23. This protein belongs to a new type of antioxidative proteins. Additionally we found that two other known proteins were induced upon exposure to oxidative stress. We also obtained a few cDNA clones that encode novel stress-induced proteins.

应激剂诱导体外培养的小鼠巨噬细胞的胱氨酸转运活性。胱氨酸进入细胞的诱导增强了谷胱甘肽（一种抗氧化细胞物质）的合成。通过差异筛选，筛选出胁迫诱导的克隆，尝试克隆胱氨酸载体。我们从小鼠巨噬细胞mRNA的1.5-3 kb片段中构建了一个定向cDNA文库，从约8 × 10^4噬菌斑中共筛选出800个克隆。将这些cDNA合成的cRNA注射非洲爪蟾卵母细胞后，我们没有检测到任何胱氨酸转运活性的表达，虽然我们不能分离编码胱氨酸载体的cNDA克隆，但我们成功地克隆了一个应激诱导的23 kDa蛋白，命名为MSP 23。该蛋白属于一种新型的抗氧化蛋白。此外，我们发现，其他两个已知的蛋白质被诱导后暴露于氧化应激。我们还获得了一些cDNA克隆，编码新的胁迫诱导蛋白。

项目成果

Sato, H., Ishii, T., Sugita, Y., Tateishi, N., and Bannai, S.: "Induction of a 23-kDa stress protein by oxidative and sulfhydryl-reactive agents in mouse peritoneal macrophages." Biochem.Biophys.Acta. 1148. 127-132 (1993)

Sato, H.、Ishii, T.、Sugita, Y.、Tateishi, N. 和 Bannai, S.：“通过小鼠腹膜巨噬细胞中的氧化和巯基反应剂诱导 23-kDa 应激蛋白。”

Miura,K.: "Cystine uptake and glutathione level in endotherial cells exposed to oxidative stress." Am.J.Physiol.262. C50-C58 (1992)

Miura,K.：“暴露于氧化应激的内皮细胞中胱氨酸的摄取和谷胱甘肽水平。”

T.Ishii: "Induction of cystine transport activity by stress" Ann.NY Acad.Sci.663. 497-498 (1992)

T.Ishii：“应激诱导胱氨酸转运活性”Ann.NY Acad.Sci.663。

H.Sato: "Induction of a 23-kDa stress protein by oxidative and sulfhydryl-reactive agents in mouse peritoneal macrophages" Biochim.Biophys.Acta. (1993)

H.Sato：“小鼠腹膜巨噬细胞中氧化和巯基反应剂诱导 23 kDa 应激蛋白”Biochim.Biophys.Acta。

H.Saito: "Expression of human intestinal dipetide transporter in Xenopus laevis oocytes" Biochem.Pharmacol.45. 776-779 (1993)

H.Saito：“非洲爪蟾卵母细胞中人肠道二肽转运蛋白的表达”Biochem.Pharmacol.45。