Defining substrate requirements for cleavage of farnesylated prelamin A by the integral membrane zinc metalloprotease ZMPSTE24.
Defining substrate requirements for cleavage of farnesylated prelamin A by the integral membrane zinc metalloprotease ZMPSTE24.
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DOI：
10.1371/journal.pone.0239269
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发表时间：
2020
期刊：
影响因子：
3.7
通讯作者：
Michaelis S
中科院分区：
文献类型：
--
作者：
Wood KM;Spear ED;Mossberg OW;Odinammadu KO;Xu W;Michaelis S
完整的膜锌金属蛋白酶ZMPSTE24在核骨架蛋白lamin A的前体--法尼化的prelamin A的蛋白分解过程中起着关键作用，这一加工步骤的失败会导致prelamin A的永久性法尼化形式的积累，从而导致Hutchinson-Gilford Progeria综合征(HGPS)以及相关的孕激素紊乱，并可能在生理性衰老中发挥作用。ZMPSTE24是一种耐人寻味的不同寻常的蛋白酶，因为它的活性部位位于一个封闭的膜内小室内，该小室由七个跨膜跨度组成，小室中有侧门允许底物进入。前层蛋白A是哺乳动物细胞中唯一已知的ZMPSTE24底物，其具体特性尚不清楚。在这项工作开始时，人们知道法尼化对于体内切割Prelamin A是必不可少的，并且Prelamin A的C-末端区域(41个氨基酸)足以识别和处理。在这里，我们使用完善的人源化酵母系统研究了ZMPSTE24切割所需的Prelamin A的其他特征。我们分析了位于ZMPSTE24裂解位点和法尼化半胱氨酸之间的Prelamin A的14个残基C-末端区域以及到裂解位点的23个残基区域，通过在Prelamin A中产生一系列丙氨酸替换、丙氨酸添加和缺失来分析Prelamin A。令人惊讶的是，我们发现对这些区域的长度和组成的特定要求有相当大的灵活性。我们讨论了这种灵活性如何与ZMPSTE24的S对Prelamin A的选择性相协调。
The integral membrane zinc metalloprotease ZMPSTE24 plays a key role in the proteolytic processing of farnesylated prelamin A, the precursor of the nuclear scaffold protein lamin A. Failure of this processing step results in the accumulation of permanently farnesylated forms of prelamin A which cause the premature aging disease Hutchinson-Gilford Progeria Syndrome (HGPS), as well as related progeroid disorders, and may also play a role in physiological aging. ZMPSTE24 is an intriguing and unusual protease because its active site is located inside of a closed intramembrane chamber formed by seven transmembrane spans with side portals in the chamber permitting substrate entry. The specific features of prelamin A that make it the sole known substrate for ZMPSTE24 in mammalian cells are not well-defined. At the outset of this work it was known that farnesylation is essential for prelamin A cleavage in vivo and that the C-terminal region of prelamin A (41 amino acids) is sufficient for recognition and processing. Here we investigated additional features of prelamin A that are required for cleavage by ZMPSTE24 using a well-established humanized yeast system. We analyzed the 14-residue C-terminal region of prelamin A that lies between the ZMPSTE24 cleavage site and the farnesylated cysteine, as well 23-residue region N-terminal to the cleavage site, by generating a series of alanine substitutions, alanine additions, and deletions in prelamin A. Surprisingly, we found that there is considerable flexibility in specific requirements for the length and composition of these regions. We discuss how this flexibility can be reconciled with ZMPSTE24’s selectivity for prelamin A.
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影响因子：
8
作者：
Clark, Kathleen M.;Jenkins, Jermaine L.;Dumont, Mark E.
通讯作者：
Dumont, Mark E.
DOI：
10.1073/pnas.192460799
发表时间：
2002-10-01
影响因子：
11.1
作者：
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通讯作者：
Young, SG
影响因子：
3.3
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影响因子：
4.8
作者：
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通讯作者：
Schmidt, Walter K.
影响因子：
11.4
作者：
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通讯作者：
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