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首页> 外文期刊>Infection and immunity >Identification and characterization of proteinase K-resistant proteins in members of the class Mollicutes.
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Identification and characterization of proteinase K-resistant proteins in members of the class Mollicutes.

机译：Mollicutes类成员中耐蛋白酶K的蛋白质的鉴定和表征。

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Proteins resistant to proteinase K are rare because of the potency, wide pH optimum, and low peptide bond specificity of this enzyme. Previously, only the prion proteins associated with transmissible spongiform encephalopathies, possibly related proteins in the mollicute Spiroplasma mirum, and proteinase K itself have been reported. We identified a new proteinase K-resistant protein, p40-pr, in two strains of Mycoplasma hyorhinis and in extracts of these organisms. p40-pr's are similar to prion proteins in their resistance to high doses of proteinase K and in the reversal of this resistance by strong denaturing conditions. However, p40-pr's were distinct immunologically, in relative molecular mass, and in their method of extraction. Two immunologically related forms of p40-pr were identified on sodium dodecyl sulfate (SDS) gels and Western immunoblots, a 40-kDa species in boiled samples and a 120-kDa species dissociable by boiling in SDS. Reduction with 2-mercaptoethanol did not affect the mass of p40-pr's or the 120-kDa forms. The development of proteinase K resistance of p40-pr correlated to age-dependent increases in organism protein-lipid ratios. p40-pr-like proteinase K-resistant proteins of 46 to 50 kDa were identified in four of eight additional species of the class Mollicutes but not in S. mirum. However, these mycoplasmal proteins did not react with antibody to the denatured 40-kDa form of M. hyorhinis p40-pr purified by electroelution. The chromatographically purified 46-kDa proteinase K-resistant protein of Mycoplasma orale was an arginine deiminase.
机译：由于这种酶的效力，最适的pH值和低的肽键特异性，因此对蛋白酶K具有抗性的蛋白很少见。以前，仅报道了与可传播的海绵状脑病相关的the病毒蛋白，可能是微小螺旋体中的相关蛋白，以及蛋白酶K本身。我们在两个支原体支原体菌株和这些生物的提取物中鉴定了一种新的耐蛋白酶K蛋白p40-pr。 p40-pr's与病毒蛋白的相似之处在于它们对高剂量蛋白酶K的抗性以及在强变性条件下的这种抗性的逆转。但是，p40-pr在免疫学，相对分子质量和提取方法上都不同。在十二烷基硫酸钠（SDS）凝胶和Western免疫印迹上鉴定出两种与免疫相关的p40-pr形式，一种是煮沸样品中的40 kDa物种，另一种是通过在SDS中煮沸可解离的120 kDa物种。用2-巯基乙醇还原不影响p40-pr或120-kDa形式的质量。 p40-pr的蛋白酶K抗性的发展与机体蛋白质-脂质比率的年龄依赖性增加有关。在Mollicutes类的8个其他物种中的4个中，发现了46至50 kDa的p40-pr样蛋白酶K抗性蛋白，而在S. mirum中没有。然而，这些支原体蛋白不与通过电洗脱纯化的变性的M. hyorhinis p40-pr的40 kDa形式的抗体反应。层析纯化的米拉支原体的46 kDa蛋白酶K抗性蛋白是精氨酸脱亚氨酶。

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