从大鼠海马cDNA文库中克隆抗n -乙酰谷氨酸肽酶。
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Bzdega T, Turi T, Wroblewska B, She D, Chung HS, Kim H, Neale JH
从大鼠海马cDNA文库中克隆抗n -乙酰谷氨酸肽酶。
神经化学杂志，1997年12月;69(6):2270-7。
- PubMed ID
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9375657 (PubMed视图］
- 摘要
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n -乙酰laspartylglutamate (NAAG)是哺乳动物神经系统中最普遍的肽神经递质。NAAG选择性地激活3型代谢性谷氨酸受体。它被神经元和神经胶质质膜胞外表面的肽酶活性灭活。编码前列腺特异性膜抗原(PSM)的人类基因已被证明能产生对抗NAAG的肽酶活性。我们克隆了人PSM cDNA，并用其构建了大鼠海马cDNA文库。我们鉴定了一个包含完整编码区域的cDNA，与PSM基因同源性为83%。预测的752个氨基酸序列与PSM序列的同源性为85%，相似度为91%。转染该cDNA的CHO细胞表达NAAG肽酶活性与从大鼠脑膜中获得的活性相似。肽酶活性被β - naag、半甘油三酯和翼氨酸抑制，但没有被天冬氨酸和翼酸抑制。原位杂交数据显示肽酶mRNA在脑内广泛分布，与肽酶活性分布一致。 The highest levels of hybridization were detected in the hippocampus, dentate gyrus, piriform cortex, choroid plexus of the ventricles, pineal gland, anterior pituitary, and supraoptic nucleus. Three transcripts (estimated at 5, 3.4, and 2.9 kb) were identified in northern blots of rat brain, while in rat kidney the third transcript appeared slightly smaller than 2.9 kb. With use of reverse transcriptase PCR with primers for the 5' end, the central region, and the 3' end of the hippocampal cDNA, the expected amplification products were obtained from rat brain RNA. Spinal cord yielded an amplification product only with primers for the 5' end of the hippocampal cDNA.