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首页> 外文期刊>Cytoskeleton >Cytosolic proteins from tobacco pollen tubes that crosslink microtubules and actin filaments in vitro are metabolic enzymes.
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Cytosolic proteins from tobacco pollen tubes that crosslink microtubules and actin filaments in vitro are metabolic enzymes.

机译：来自烟草花粉管的在体外交联微管和肌动蛋白丝的胞质蛋白是代谢酶。

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摘要

In plant cells, many processes require cooperative action of both microtubules and actin filaments, but proteins mediating interactions between these cytoskeletal members are mostly undiscovered. Here, we attempt to identify such proteins by affinity purification. Cytosol from Nicotiana tabacum (tobacco) pollen tubes was incubated first with actin filaments, and then proteins eluted from the actin were incubated with microtubules, and finally those microtubule-binding proteins were pooled in an active fraction. This fraction bundled actin filaments but not microtubules. However, when the fraction was added to both actin and microtubules, large bundles resulted, containing both polymers, regardless of the order of addition of components. Similar results were obtained when the order of affinity purification was reversed. The four most abundant bands from the fractions were identified from peptide fragments analyzed by mass spectrometry. The same four proteins were identified regardless of the order of affinity purification. The proteins are: homocysteine methyltransferase, phosphofructokinase, pyruvate decarboxylase, and glucan protein synthase (reversibly glycosylated protein). These results suggest the importance of structuring metabolism within the confines of the pollen tube cytoplasm.
机译：在植物细胞中，许多过程都需要微管和肌动蛋白丝的协同作用，但是介导这些细胞骨架成员之间相互作用的蛋白质却大多未被发现。在这里，我们尝试通过亲和纯化鉴定此类蛋白质。首先将烟草花粉管中的细胞溶胶与肌动蛋白丝一起孵育，然后从肌动蛋白洗脱的蛋白质与微管一起孵育，最后将这些微管结合蛋白合并到一个活性级分中。该级分捆绑肌动蛋白丝，但不捆绑微管。但是，当将级分同时添加到肌动蛋白和微管中时，无论组分的添加顺序如何，都会产生包含两种聚合物的大束。当亲和纯化的顺序颠倒时，可获得相似的结果。通过质谱分析从肽片段中鉴定出馏分中四个最丰富的条带。无论亲和纯化的顺序如何，都可以鉴定出相同的四种蛋白质。蛋白质是：高半胱氨酸甲基转移酶，磷酸果糖激酶，丙酮酸脱羧酶和葡聚糖蛋白质合酶（可逆糖基化蛋白质）。这些结果表明在花粉管细胞质范围内构造代谢的重要性。

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