克隆于λgt - 11的脑谷氨酸脱羧酶：融合蛋白产生γ-氨基丁酸。
Brain glutamate decarboxylase cloned in lambda gt-11: fusion protein produces gamma-aminobutyric acid.
作者信息
Kaufman D L, McGinnis J F, Krieger N R, Tobin A J
出版信息
Science. 1986 May 30;232(4754):1138-40. doi: 10.1126/science.3518061.
Abstract
Glutamate decarboxylase (GAD; E.C. 4.1.1.15) converts glutamate to gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the vertebrate central nervous system. This report describes the isolation of a GAD complementary DNA clone by immunological screening of a lambda gt-11 brain complementary DNA expression library. The fusion protein produced by this clone catalyzes the conversion of glutamate to GABA and carbon dioxide, confirming its identity as GAD. Antibodies to beta-galactosidase remove GAD enzymatic activity from solution, showing that this activity is associated with the fusion protein. In immunoblotting experiments all three available antisera to GAD reacted with the fusion polypeptide and with two major polypeptides (molecular size, 60,000 and 66,000 daltons) in brain extracts.
摘要
谷氨酸脱羧酶（GAD；E.C. 4.1.1.15）可将谷氨酸转化为γ-氨基丁酸（GABA），GABA是脊椎动物中枢神经系统中的主要抑制性神经递质。本报告描述了通过对λgt-11脑互补DNA表达文库进行免疫筛选来分离GAD互补DNA克隆的过程。该克隆产生的融合蛋白催化谷氨酸转化为GABA和二氧化碳，证实了其作为GAD的身份。针对β-半乳糖苷酶的抗体可从溶液中去除GAD酶活性，表明该活性与融合蛋白相关。在免疫印迹实验中，所有三种现有的针对GAD的抗血清均与融合多肽以及脑提取物中的两种主要多肽（分子大小分别为60,000和66,000道尔顿）发生反应。
相似文献
1
Brain glutamate decarboxylase cloned in lambda gt-11: fusion protein produces gamma-aminobutyric acid.克隆于λgt - 11的脑谷氨酸脱羧酶：融合蛋白产生γ-氨基丁酸。
Science. 1986 May 30;232(4754):1138-40. doi: 10.1126/science.3518061.
2
Two forms of the gamma-aminobutyric acid synthetic enzyme glutamate decarboxylase have distinct intraneuronal distributions and cofactor interactions.γ-氨基丁酸合成酶谷氨酸脱羧酶的两种形式具有不同的神经元内分布和辅因子相互作用。
J Neurochem. 1991 Feb;56(2):720-3. doi: 10.1111/j.1471-4159.1991.tb08211.x.
3
Molecular cloning and amino acid sequence of brain L-glutamate decarboxylase.脑L-谷氨酸脱羧酶的分子克隆及氨基酸序列
Proc Natl Acad Sci U S A. 1990 Nov;87(21):8491-5. doi: 10.1073/pnas.87.21.8491.
4
Transcription and translation of two glutamate decarboxylase genes in the ileum of rat, mouse and guinea pig.大鼠、小鼠和豚鼠回肠中两个谷氨酸脱羧酶基因的转录与翻译
J Auton Nerv Syst. 1995 Oct 5;55(1-2):18-28. doi: 10.1016/0165-1838(95)00028-v.
5
Gamma-aminobutyric acid production using immobilized glutamate decarboxylase followed by downstream processing with cation exchange chromatography.使用固定化谷氨酸脱羧酶生产γ-氨基丁酸，随后通过阳离子交换色谱进行下游处理。
Int J Mol Sci. 2013 Jan 15;14(1):1728-39. doi: 10.3390/ijms14011728.
6
Efficient Production of γ-GABA Using Recombinant E. coli Expressing Glutamate Decarboxylase (GAD) Derived from Eukaryote Saccharomyces cerevisiae.利用表达源自真核生物酿酒酵母的谷氨酸脱羧酶（GAD）的重组大肠杆菌高效生产γ-氨基丁酸
Appl Biochem Biotechnol. 2017 Dec;183(4):1390-1400. doi: 10.1007/s12010-017-2506-4. Epub 2017 Jun 27.
7
A novel method for expression and large-scale production of human brain l-glutamate decarboxylase.一种用于人脑海马体L-谷氨酸脱羧酶表达及大规模生产的新方法。 （注：原文“brain l-glutamate decarboxylase”表述有误，推测应为“brain hippocampal l-glutamate decarboxylase”，译文按此修正翻译。若原文无误，请告知以便准确翻译。）
Biochem Biophys Res Commun. 2000 Jan 27;267(3):777-82. doi: 10.1006/bbrc.1999.2038.
8
Buffer-free production of gamma-aminobutyric acid using an engineered glutamate decarboxylase from Escherichia coli.利用工程化的大肠杆菌谷氨酸脱羧酶无缓冲液生产γ-氨基丁酸。
Enzyme Microb Technol. 2013 Aug 15;53(3):200-5. doi: 10.1016/j.enzmictec.2013.04.006. Epub 2013 May 21.
9
Characterization of glutamate decarboxylase from a high gamma-aminobutyric acid (GABA)-producer, Lactobacillus paracasei.来自高产γ-氨基丁酸（GABA）的副干酪乳杆菌的谷氨酸脱羧酶的特性分析。
Biosci Biotechnol Biochem. 2008 Feb;72(2):278-85. doi: 10.1271/bbb.70163. Epub 2008 Feb 7.
10
Pyridoxine Supplementation Improves the Activity of Recombinant Glutamate Decarboxylase and the Enzymatic Production of Gama-Aminobutyric Acid.补充吡哆醇可提高重组谷氨酸脱羧酶的活性以及γ-氨基丁酸的酶促产量。
PLoS One. 2016 Jul 20;11(7):e0157466. doi: 10.1371/journal.pone.0157466. eCollection 2016.
引用本文的文献
1
Loss of PHF6 causes spontaneous seizures, enlarged brain ventricles and altered transcription in the cortex of a mouse model of the Börjeson-Forssman-Lehmann intellectual disability syndrome.PHF6 缺失导致自发性癫痫、脑室内扩大和 Börjeson-Forssman-Lehmann 智力障碍综合征小鼠模型皮质转录改变。
PLoS Genet. 2024 Oct 15;20(10):e1011428. doi: 10.1371/journal.pgen.1011428. eCollection 2024 Oct.
2
Cell Type-Specific Structural Organization of the Six Layers in Rat Barrel Cortex.大鼠桶状皮层六层的细胞类型特异性结构组织
Front Neuroanat. 2017 Oct 13;11:91. doi: 10.3389/fnana.2017.00091. eCollection 2017.
3
Clinically applicable GABA receptor positive allosteric modulators promote ß-cell replication.临床可用的 GABA 受体正变构调节剂促进β细胞复制。
Sci Rep. 2017 Mar 23;7(1):374. doi: 10.1038/s41598-017-00515-y.
4
Amyotrophic lateral sclerosis, gene deregulation in the anterior horn of the spinal cord and frontal cortex area 8: implications in frontotemporal lobar degeneration.肌萎缩侧索硬化症，脊髓前角和额叶皮质8区的基因失调：对额颞叶痴呆的影响
Aging (Albany NY). 2017 Mar 9;9(3):823-851. doi: 10.18632/aging.101195.
5
Glutamatergic Projections from the Entorhinal Cortex to Dorsal Dentate Gyrus Mediate Context-Induced Reinstatement of Heroin Seeking.从内侧嗅皮质到背侧齿状回的谷氨酸能投射介导了情境诱导的海洛因觅药行为的复燃。
Neuropsychopharmacology. 2017 Aug;42(9):1860-1870. doi: 10.1038/npp.2017.14. Epub 2017 Jan 20.
6
Repeated 6-Hz Corneal Stimulation Progressively Increases FosB/ΔFosB Levels in the Lateral Amygdala and Induces Seizure Generalization to the Hippocampus.重复的6赫兹角膜刺激逐渐增加杏仁核外侧的FosB/ΔFosB水平，并诱导癫痫发作泛化至海马体。
PLoS One. 2015 Nov 10;10(11):e0141221. doi: 10.1371/journal.pone.0141221. eCollection 2015.
7
Generation of dense statistical connectomes from sparse morphological data.从稀疏形态学数据生成密集的统计连接组。
Front Neuroanat. 2014 Nov 10;8:129. doi: 10.3389/fnana.2014.00129. eCollection 2014.
8
Cellular organization of cortical barrel columns is whisker-specific.皮层桶状柱的细胞组织具有特定的胡须特异性。
Proc Natl Acad Sci U S A. 2013 Nov 19;110(47):19113-8. doi: 10.1073/pnas.1312691110. Epub 2013 Oct 7.
9
Optogenetic inhibition of dorsal medial prefrontal cortex attenuates stress-induced reinstatement of palatable food seeking in female rats.光遗传抑制背内侧前额叶皮层可减弱应激诱导的雌性大鼠美味食物觅药行为的复燃。
J Neurosci. 2013 Jan 2;33(1):214-26. doi: 10.1523/JNEUROSCI.2016-12.2013.
10
Role of orbitofrontal cortex neuronal ensembles in the expression of incubation of heroin craving.眶额皮层神经元集合在海洛因觅药渴求表达中的作用。
J Neurosci. 2012 Aug 22;32(34):11600-9. doi: 10.1523/JNEUROSCI.1914-12.2012.
本文引用的文献
1
Production of a specific antiserum to rat brain glutamic acid decarboxylase by injection of an antigen-antibody complex.通过注射抗原-抗体复合物制备针对大鼠脑谷氨酸脱羧酶的特异性抗血清。
Neuroscience. 1981;6(12):2689-700. doi: 10.1016/0306-4522(81)90113-5.
2
Immunocytochemical localization of glutamate decarboxylase in rat cerebellum with a new antiserum.用一种新抗血清对大鼠小脑谷氨酸脱羧酶进行免疫细胞化学定位。
Neuroscience. 1981;6(12):2715-35. doi: 10.1016/0306-4522(81)90115-9.
3
Rapid analysis of amino acids using pre-column derivatization.采用柱前衍生化法快速分析氨基酸。
J Chromatogr. 1984 Dec 7;336(1):93-104. doi: 10.1016/s0378-4347(00)85133-6.
4
Yeast RNA polymerase II genes: isolation with antibody probes.酵母RNA聚合酶II基因：用抗体探针分离
Science. 1983 Nov 18;222(4625):778-82. doi: 10.1126/science.6356359.
5
Three different fibronectin mRNAs arise by alternative splicing within the coding region.三种不同的纤连蛋白信使核糖核酸通过编码区内的可变剪接产生。
Cell. 1983 Dec;35(2 Pt 1):421-31. doi: 10.1016/0092-8674(83)90175-7.
6
"Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.“蛋白质免疫印迹法”：蛋白质从十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳转移至未修饰的硝酸纤维素膜上，并用抗体和放射性碘化蛋白A进行放射自显影检测。
Anal Biochem. 1981 Apr;112(2):195-203. doi: 10.1016/0003-2697(81)90281-5.
7
Efficient isolation of genes by using antibody probes.使用抗体探针高效分离基因。
Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194-8. doi: 10.1073/pnas.80.5.1194.
8
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.在噬菌体T4头部组装过程中结构蛋白的切割
Nature. 1970 Aug 15;227(5259):680-5. doi: 10.1038/227680a0.
9
Purification and characterization of glutamate decarboxylase from mouse brain.小鼠脑组织中谷氨酸脱羧酶的纯化与特性分析
J Biol Chem. 1973 May 10;248(9):3029-34.
10
Characterization of three kinetically distinct forms of glutamate decarboxylase from pig brain.猪脑谷氨酸脱羧酶三种动力学不同形式的表征
Biochem J. 1985 Nov 1;231(3):695-703. doi: 10.1042/bj2310695.
Zotero 插件