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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >MOLECULAR CLONING AND FUNCTIONAL CHARACTERIZATION OF THE XENOPUS CA2+-BINDING PROTEIN FREQUENIN
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MOLECULAR CLONING AND FUNCTIONAL CHARACTERIZATION OF THE XENOPUS CA2+-BINDING PROTEIN FREQUENIN

机译：Xenopus CA2 +结合蛋白频率的分子克隆和功能表征

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摘要

Frequenin was originally identified in Drosophila melanogaster as a Ca2+-binding protein facilitating transmitter release at the neuromuscular junction. We have cloned the Xenopus frequenin (Xfreq) by PCR using degenerate primers combined with low-stringency hybridization. The deduced protein has 70% identity with Drosophila frequenin and about 38-58% identity with other Ca2+-binding proteins. The most prominent features ari: the four EF-hands, Ca2+-binding motifs. Xfreq mRNA is abundant in the brain and virtually nondetectable from adult muscle. Western blot analysis indicated that Xfreq is highly concentrated in the adult brain and is absent from nonneural tissues such as heart and kidney. During development, the expression of the protein correlated well with the maturation of neuromuscular synapses. To determine the function of Xfreq at the developing neuromuscular junction, the recombinant protein was introduced into Xenopus embryonic spinal neurons by early blastomere injection. Synapses made by spinal neurons containing exogenous Xfreq exhibited a much higher synaptic efficacy. These results provide direct evidence that frequenin enhances transmitter release at the vertebrate neuromuscular synapse and suggest its potential role in synaptic development and plasticity. [References: 21]
机译：Frequenin最初在果蝇中被鉴定为Ca2 +结合蛋白，可促进神经肌肉接头处的递质释放。我们已经通过简并引物结合低严格性杂交通过PCR克隆了非洲爪蟾frequenin（Xfreq）。推导的蛋白与果蝇frequenin具有70％的同一性，与其他Ca2 +结合蛋白具有约38-58％的同一性。 ari的最突出特征是：四个EF手，Ca2 +结合基序。 Xfreq mRNA在大脑中很丰富，几乎无法从成年肌肉中检测到。蛋白质印迹分析表明Xfreq高度集中在成年大脑中，而非神经组织（如心脏和肾脏）中则不存在。在发育过程中，蛋白质的表达与神经肌肉突触的成熟密切相关。为了确定Xfreq在发育中的神经肌肉连接处的功能，通过早期卵裂球注射将重组蛋白引入非洲爪蟾胚胎脊髓神经元。含有外源Xfreq的脊髓神经元产生的突触表现出更高的突触功效。这些结果提供了直接的证据，表明frequenin增强了脊椎动物神经肌肉突触中的递质释放，并暗示了其在突触发育和可塑性中的潜在作用。 [参考：21]

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