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Changing pattern of cyclic AMP-binding proteins during germination of Mucor racemosus sporangiospores.

机译:环毛孢子孢子萌发过程中环状AMP结合蛋白的变化模式。

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摘要

Interation of cyclic AMP with a profoundly changing pattern of specific binding proteins was shown during aerobic germination of sporangiospores from the fungus Mucor racemosus. 32P-labeled 8-azido-cycli AMP, an analogue of cyclic AMP that forms a covalent linkage with the binding proteins under u.v. light, was used as the ligand. Binding proteins carrying this photoaffinity label were separated by polyacrylamide-gel electrophoresis and identified by radioautography. Equibiltrium dissociation constants (Kd) and binding-response curves in the presence of competing nucleotides were identical for both 8-azido-cyclic [32P]AMP and cyclic [3H]AMP. A quantitative binding assay with both 8-azido-cyclic [32P]AMP and cyclic [3H]AMP over the time course of sporangiospore germination indicated a parallel relationship between cyclic AMP-binding capacity and the intracellular concentrations of cyclic AMP reported in a previous study [Paznokas & Sypherd (1975) J. Bacteriol. 124, 134--139]. Both of these parameters attained transient high values at a time of development when addition of exogenous cyclic AMP prevents hyphal-germ-tube emergence. The measured Kd values did not change during sport germination.
机译:在真菌Mucor racemosus的孢子孢子有氧萌发过程中,环状AMP与特异结合蛋白的模式发生了深刻变化。 32P标记的8-叠氮基-环AMP,在uv下与结合蛋白形成共价键的环状AMP的类似物。光,用作配体。带有该光亲和标记的结合蛋白通过聚丙烯酰胺凝胶电泳分离,并通过放射自显影进行鉴定。对于8-叠氮基环[32P] AMP和环[3H] AMP而言,在竞争核苷酸存在下的等价解离常数(Kd)和结合反应曲线是相同的。在孢子囊孢子萌发的过程中,同时使用8-叠氮基环[32P] AMP和环[3H] AMP进行定量结合测定表明,先前研究中报道的环AMP结合能力与环内AMP的细胞内浓度之间存在平行关系[Paznokas&Sypherd(1975)J. Bacteriol。 124,134--139]。当添加外源性环状AMP阻止菌丝-细菌管出现时,这两个参数在开发时均达到瞬时高值。在运动萌发过程中,测得的Kd值不变。

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  • 作者

    Orlowski, M;

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  • 年度 1979
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  • 原文格式 PDF
  • 正文语种 en
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