Proteolytic Cleavage of Staphylococcal Exoproteins Analyzed by Two‐Dimensional Gel Electrophoresis
Proteolytic Cleavage of Staphylococcal Exoproteins Analyzed by Two‐Dimensional Gel Electrophoresis
复制标题
二维凝胶电泳分析葡萄球菌外蛋白的蛋白水解切割
作者：
Y. Kawano;Mika Kawagishi;M. Nakano;Kiyomi Mase;T. Yamashino;T. Hasegawa;M. Ohta
金黄色葡萄球菌的胞外蛋白酶是一种与金黄色葡萄球菌感染致病性相关的潜在毒力因子。这些蛋白酶也可能参与从该生物体释放的其他外蛋白的蛋白水解切割。为了确定目标外蛋白及其被蛋白酶切割的位点，进行了高分辨率二维聚丙烯酰胺凝胶电泳，然后进行了外蛋白点的N末端氨基酸测序。在S.金黄色葡萄球菌NCTC 8325，然后在后期稳定阶段，由于进一步的蛋白水解降解，大多数这些高分子蛋白质斑点变得不可见。作为N末端分析的结果，在早期稳定期检测到脂肪酶、三酰甘油脂肪酶、orf 619蛋白和orf 388蛋白作为多个斑点。我们发现这些外蛋白分别在3、7、4和4个不同的位点被蛋白酶切割。根据M.W.从凝胶中获得的每个肽点的pI和pI，以及它们与计算值的匹配以及它们的N末端序列，我们表明推定肽的位置是由这些蛋白质的蛋白水解切割产生的。
Extracellular proteases of Staphylococcus aureus are emerging as potential virulence factors that are relevant to the pathogenicity of staphylococcal infections. These proteases may also be involved in the proteolytic cleavage of other exoproteins released from this organism. To define the target exoproteins and their sites of cleavage by proteases, high‐resolution two‐dimensional polyacrylamide gel electrophoresis followed by N‐terminal amino acid sequencing of exoprotein spots was performed. Two to three hundred exoprotein spots were detected at the early‐stationary phase of cultures of S. aureus NCTC8325, and then at the late‐stationary stage most of these high molecular protein spots became invisible due to further proteolytic degradation. As the result of N‐terminal analysis, lipase, triacylglycerol lipase, orf619 protein and orf388 protein were detected as multiple spots at the early‐stationary phase. We found that these exoproteins were cleaved at 3, 7, 4 and 4 different sites, respectively, by proteases. According to the M.W. and pI of each peptide spot obtained from the gel and their matches with calculated values in addition to their N‐terminal sequences, we showed that the positions of putative peptides resulted from proteolytic cleavage of these proteins.
DOI：
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发表时间：
1995-03
期刊：
Infectious agents and disease
影响因子：
--
作者：
J. Goguen;N. Hoe;Y. Subrahmanyam
通讯作者：
J. Goguen;N. Hoe;Y. Subrahmanyam
金黄色葡萄球菌胞外毒素的遗传分析。
影响因子：
10.5
作者：
Iandolo,JJ
通讯作者：
Iandolo,JJ