Characterization of an Extracellular Serine Protease of Fusarium eumartii and its Action on Pathogenesis Related Proteins
Characterization of an Extracellular Serine Protease of Fusarium eumartii and its Action on Pathogenesis Related Proteins
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真镰刀菌胞外丝氨酸蛋白酶的表征及其对发病机制相关蛋白的作用
DOI：
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发表时间：
2004
影响因子：
1.8
通讯作者：
C. Casalongué
中科院分区：
文献类型：
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作者：
F. Olivieri;María Eugenia Zanetti;C. Oliva;A. Covarrubias;C. Casalongué
蛋白酶已被提议作为某些病原真菌入侵策略的一部分。在这项工作中，由植物病原真菌Fusarium solani f.sp.产生的丝氨酸蛋白酶。 eumartii 被纯化并表征。通过 Superose 12 柱进行凝胶过滤，然后在 Phenyl Superose 中进行疏水相互作用层析，并通过 Superdex 75 进行凝胶过滤层析，从而完成酶的纯化。通过 SDS/PAGE（未经热处理）对纯化的酶进行分析，显示出一条单条带，这对应于通过酶谱检测到的蛋白水解活性。当该蛋白质经受变性条件时，显露出约 30 和 33kDa 的两种主要多肽。这些多肽之一的N端氨基酸序列显示与枯草杆菌蛋白酶家族的真菌成熟丝氨酸蛋白酶高度相似。该蛋白酶在体外水解马铃薯细胞间洗涤液和细胞壁的特定多肽。该蛋白酶还能够降解细胞间洗涤液中与发病机制相关的蛋白质。讨论了这种丝氨酸蛋白酶作为真菌定殖马铃薯块茎组织策略的一部分的作用。
Proteases have been proposed as part of the invasion strategies of some pathogenic fungi. In this work, a serine protease produced by the phytopathogenic fungus Fusarium solani f.sp. eumartii was purified and characterized. Purification of the enzyme was accomplished by gel filtration through a Superose 12 column, followed by hydrophobic interaction chromatography in Phenyl Superose and gel filtration chromatography through Superdex 75. Analysis of the purified enzyme by SDS/PAGE without heat treatment, revealed a single band, which corresponded to the proteolytic activity detected by zymogram. When this protein was subjected to denaturing conditions, two major polypeptides of approximately 30 and 33kDa were revealed. The N-terminal amino acid sequence of one of these polypeptides showed a high similarity with fungal mature serine proteases of the subtilisin family. This protease hydrolysed in vitro, specific polypeptides of potato intercellular washing fluids and cell walls. The protease was also able to degrade pathogenesis-related proteins from the intercellular washing fluids. The role of this serine protease as part of the fungal strategy to colonize potato tuber tissues is discussed.
来自 Cochliobolus Carbonum 的三种胞外蛋白酶：ALP1 的克隆和靶向破坏。
DOI：
10.1094/mpmi-9-0290
发表时间：
1996
期刊：
Molecular plant-microbe interactions : MPMI.
影响因子：
--
作者：
Murphy,JM;Walton,JD
通讯作者：
Walton,JD