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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Posttranslational modification of G alpha o 1 generates G alpha o 3, an abundant G protein in brain
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Posttranslational modification of G alpha o 1 generates G alpha o 3, an abundant G protein in brain

机译：G alpha o 1的翻译后修饰产生G alpha o 3，这是大脑中丰富的G蛋白

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摘要

G ALPHA O , the most abundant G protein in mam- malian brain, occurs at least in two subforms, i.e., G alpha o 1 and G alpha o 2, derived by alternative splicing of the mRNA. A third G alpha o 1-related isoform, G alpha o3, has been purified, representing about 30 of total Go in brain. Initial studies rcvealed distinct biochemical properties of G alpha o 3 as compared with other G alpha o isoforms. In matrix-assisted laser desorption/ionization pep- tide mass mapping of gel-isolatcd G alpha o 1 and Galpha o 3, C-terminal peptides showed a difference of +l Da for G alpha o3. Nanoclec- trospray tandem mass spectrometry sequencing revealed an Asp instead of an Asn at position 346 of G alpha o 3. Gel clectro. phoretic analysis of recombinanl G alpha o3 showed the same mobility as native G alpha o3 but distinct to G alpha o 1. The conversion of 346Asn -> Asp changed the signaling properties, including the velocity of the basal guanine nucleotide-exchange reaction, which points to the involvement of the C terminus in basal guanosine 5'- [r-thio] triphosphate binding. No cDNA coding for G alphaa o 3 was detectcd, suggesting an enzymatic deamidation of G alpha o1 by a yet-unidentified activity. Therefore, G alpha hetero- geneity is generated not only at the DNA or RNA levels, but also at the protein level. The relative amount of G alpha o1 and G alpha o 3 different from cell type to cell type, indicating an additional principle of G protein regulation.
机译：哺乳动物脑中最丰富的G蛋白G ALPHA O至少以两种亚型出现，即通过交替剪接mRNA衍生而来的G alpha o 1和G alpha o 2。已纯化出第三个与G alpha o 1相关的同种型，G alpha o3，约占大脑总Go的30。初步研究显示，与其他G alpha o同工型相比，G alpha o 3具有独特的生化特性。在凝胶分离的G alpha o 1和Galpha o 3的基质辅助激光解吸/电离肽质量质谱图中，C末端肽对G alpha o3的差异为+1 Da。纳米喷雾串联质谱分析显示在Gαo 3的346位为Asp而不是Asn。重组Gαo3的电泳分析显示出与天然G alpha o3相同的迁移率，但与G alpha o 1不同。346Asn-> Asp的转化改变了信号传导特性，包括基础鸟嘌呤核苷酸交换反应的速度，这表明C端参与基础鸟苷5'-[r-硫代]三磷酸结合。没有检测到编码G alphaa o 3的cDNA，这表明G alpha o1的酶促脱酰胺作用尚不明确。因此，不仅在DNA或RNA水平，而且在蛋白质水平都产生G alpha异质性。不同细胞类型的G alpha o1和G alpha o 3的相对量不同，这表明了G蛋白调节的另一原理。

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