The Final Maturation State of β-actin Involves N-terminal Acetylation by NAA80, not N-terminal Arginylation by ATE1.
The Final Maturation State of β-actin Involves N-terminal Acetylation by NAA80, not N-terminal Arginylation by ATE1.
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DOI：
10.1016/j.jmb.2021.167397
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发表时间：
2022-01-30
影响因子：
5.6
通讯作者：
Arnesen T
中科院分区：
文献类型：
--
作者：
Drazic A;Timmerman E;Kajan U;Marie M;Varland S;Impens F;Gevaert K;Arnesen T
肌动蛋白是真核细胞中细胞骨架的标志性蛋白，影响一系列细胞功能。肌动蛋白动力学通过无数肌动蛋白结合蛋白和翻译后修饰来调节。哺乳动物肌动蛋白家族由六种不同的同种型组成，它们的N-末端（Nt）序列略有不同。在合成期间和合成之后，肌动蛋白经历复杂的NT加工，产生成熟的肌动蛋白亚型。广泛表达的细胞质β-肌动蛋白被N-α乙酰转移酶80（NAA 80）NT-乙酰化，产生NT-序列Ac-DDDI-。此外，β-actin也被报道在进一步的肽酶介导的加工后被乙酰转移酶1（ATE 1）Nt-actin化，产生RDDI-。为了详细描述肌动蛋白的NT加工，我们使用了最先进的蛋白质组学。为了估计相对细胞水平的NT-修饰的肌动蛋白的proteoforms，我们采用野生型和NAA 80缺乏细胞，其中肌动蛋白是没有NT-乙酰化。我们发现，靶向蛋白质组学是上级的商业上可获得的抗体，以前用于分析β-肌动蛋白的NT-β化。值得注意的是，尽管使用了灵敏的基于质谱的技术，但我们无法证实先前声称的NT-乙酰化β-肌动蛋白（RDDI-）在野生型或缺乏NAA 80的细胞中的存在。可以识别出最初切割的β-肌动蛋白（DDDI-）的极少量Nt-arginylation，但仅在缺乏NAA 80的细胞中，而在野生型细胞中则不然。我们还鉴定了小部分切割和未修饰的β-肌动蛋白（DDI-）以及切割和N-乙酰化的β-肌动蛋白（Ac-DDI-）。总之，我们表明β-肌动蛋白的多步NT-成熟被NAA 80终止，其NT-乙酰化暴露的Nt-Asp残基，实际上不存在先前声称的NT-乙酰化。
Actin is a hallmark protein of the cytoskeleton in eukaryotic cells, affecting a range of cellular functions. Actin dynamics is regulated through a myriad of actin-binding proteins and post-translational modifications. The mammalian actin family consists of six different isoforms, which vary slightly in their N-terminal (Nt) sequences. During and after synthesis, actins undergo an intricate Nt-processing that yields mature actin isoforms. The ubiquitously expressed cytoplasmic β-actin is Nt-acetylated by N-alpha acetyltransferase 80 (NAA80) yielding the Nt-sequence Ac-DDDI-. In addition, β-actin was also reported to be Nt-arginylated by arginyltransferase 1 (ATE1) after further peptidase-mediated processing, yielding RDDI-. To characterize in detail the Nt-processing of actin, we used state-of-the-art proteomics. To estimate the relative cellular levels of Nt-modified proteoforms of actin, we employed wildtype and NAA80-lacking cells, in which actin was not Nt-acetylated. We found that targeted proteomics is superior to a commercially available antibody previously used to analyze Nt-arginylation of β-actin. Significantly, despite the use of sensitive mass spectrometry-based techniques, we could not confirm the existence of the previously claimed Nt-arginylated β-actin (RDDI-) in either wildtype or NAA80-lacking cells. A very minor level of Nt-arginylation of the initially cleaved β-actin (DDDI-) could be identified, but only in NAA80-lacking cells, not in wildtype cells. We also identified small fractions of cleaved and unmodified β-actin (DDI-) as well as cleaved and Nt-acetylated β-actin (Ac-DDI-). In sum, we show that the multi-step Nt-maturation of β-actin is terminated by NAA80, which Nt-acetylates the exposed Nt-Asp residues, in the virtual absence of previously claimed Nt-arginylation.
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DOI：
10.1007/978-1-4939-6850-3_2
发表时间：
2017-01-01
期刊：
PROTEIN TERMINAL PROFILING: METHODS AND PROTOCOLS
影响因子：
--
作者：
Foyn, Havard;Thompson, Paul R.;Arnesen, Thomas
通讯作者：
Arnesen, Thomas
DOI：
10.1073/pnas.1718336115
发表时间：
2018-04-24
影响因子：
11.1
作者：
Drazic A;Aksnes H;Marie M;Boczkowska M;Varland S;Timmerman E;Foyn H;Glomnes N;Rebowski G;Impens F;Gevaert K;Dominguez R;Arnesen T
通讯作者：
Arnesen T
影响因子：
2.4
作者：
Beigl TB;Kjosås I;Seljeseth E;Glomnes N;Aksnes H
通讯作者：
Aksnes H
影响因子：
21.3
作者：
Cha-Molstad, Hyunjoo;Sung, Ki Sa;Hwang, Joonsung;Kim, Kyoung A.;Yu, Ji Eun;Yoo, Young Dong;Jang, Jun Min;Han, Dong Hoon;Molstad, Michael;Kim, Jung Gi;Lee, Yoon Jee;Zakrzewska, Adriana;Kim, Su-Hyeon;Kim, Sung Tae;Kim, Sun Yong;Lee, Hee Gu;Soung, Nak Kyun;Ahn, Jong Seog;Ciechanover, Aaron;Kim, Bo Yeon;Kwon, Yong Tae
通讯作者：
Kwon, Yong Tae
影响因子：
4.8
作者：
Eisenach, Patricia A.;Schikora, Franziska;Posern, Guido
通讯作者：
Posern, Guido