不同蛋白酶体亚群的生化纯化。

Enzyme & protein Pub Date : 1993-01-01 DOI:10.1159/000468692
M G Brown, J J Monaco
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引用次数: 18

摘要

蛋白酶体是复杂的细胞蛋白酶,能够在体外降解许多蛋白质和肽底物。这些粒子结构复杂;它们由至少14个小分子质量的多肽亚基组装而成成熟的20S蛋白酶体。最近,我们证明蛋白酶体亚群可以通过血清学标准进行区分,并发现蛋白酶体亚基组成的细微差异可以改变其切割特异性。当一些蛋白酶体与其他蛋白质结合形成26S ATP和泛素依赖性蛋白酶时,蛋白酶体结构的复杂性进一步增强。在此,我们证实了不同细胞形式的蛋白酶体的存在,并表明它们在疏水特性上有所不同。我们使用单独的生化技术可重复纯化不同的蛋白酶体亚群,类似于血清学定义的LMP2+和LMP2-蛋白酶体。与血清学定义的LMP2+和LMP2-蛋白酶体相比,这些蛋白酶体亚群在至少三种多肽的表达上有所不同,并且都缺乏一些额外的多肽。最后,我们证明了这些结构独特的蛋白酶体在切割特定的荧光肽底物方面的能力不同。哺乳动物细胞可能招募和修饰蛋白酶体来执行不同的细胞任务。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Biochemical purification of distinct proteasome subsets.

Proteasomes are intricate cellular proteases that are able to degrade many protein and peptide substrates in vitro. These particles are structurally complex; they are assembled from at least 14 small molecular mass polypeptide subunits to form mature 20S proteasomes. Recently, we demonstrated that proteasome subsets may be discriminated by serological criteria, and have found that subtle differences in the subunit composition of proteasomes can alter their cleavage specificity. Proteasome structural complexity is further enhanced when some proteasomes associate with additional proteins to form a 26S ATP- and ubiquitin-dependent protease. Herein we confirm the existence of distinct cellular forms of proteasomes, and show that they differ in their hydrophobic characteristics. We have reproducibly purified, using solely biochemical techniques, distinct proteasome subsets similar to the serologically defined LMP2+ and LMP2- proteasomes. These proteasome subsets differ in their expression of at least three polypeptides, and both lack several additional polypeptides as compared to the serologically defined LMP2+ and LMP2- proteasomes. Finally, we demonstrate that these structurally unique proteasomes differ in their capacity to cleave a defined panel of fluorogenic peptide substrates. It appears that mammalian cells might recruit and modify proteasomes to perform distinct cellular tasks.

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