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Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10

Arrest defective 1 (ARD1), also known as N(alpha)-acetyltransferase 10 (NAA10) was originally identified as an N-terminal acetyltransferase (NAT) that catalyzes the acetylation of N-termini of newly synthesized peptides. After that, mammalian ARD1/NAA10 expanded its’ role to lysine acetyltransferase...

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Autores principales: Vo, Tam Thuy Lu, Park, Ji-Hyeon, Lee, Eun Ji, Nguyen, Yen Thi Kim, Han, Byung Woo, Nguyen, Hien Thi Thu, Mun, Kyo Cheol, Ha, Eunyoung, Kwon, Taeg Kyu, Kim, Kyu-Won, Jeong, Chul-Ho, Seo, Ji Hae
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7036845/
https://www.ncbi.nlm.nih.gov/pubmed/32013195
http://dx.doi.org/10.3390/molecules25030588
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author Vo, Tam Thuy Lu
Park, Ji-Hyeon
Lee, Eun Ji
Nguyen, Yen Thi Kim
Han, Byung Woo
Nguyen, Hien Thi Thu
Mun, Kyo Cheol
Ha, Eunyoung
Kwon, Taeg Kyu
Kim, Kyu-Won
Jeong, Chul-Ho
Seo, Ji Hae
author_facet Vo, Tam Thuy Lu
Park, Ji-Hyeon
Lee, Eun Ji
Nguyen, Yen Thi Kim
Han, Byung Woo
Nguyen, Hien Thi Thu
Mun, Kyo Cheol
Ha, Eunyoung
Kwon, Taeg Kyu
Kim, Kyu-Won
Jeong, Chul-Ho
Seo, Ji Hae
author_sort Vo, Tam Thuy Lu
collection PubMed
description Arrest defective 1 (ARD1), also known as N(alpha)-acetyltransferase 10 (NAA10) was originally identified as an N-terminal acetyltransferase (NAT) that catalyzes the acetylation of N-termini of newly synthesized peptides. After that, mammalian ARD1/NAA10 expanded its’ role to lysine acetyltransferase (KAT) that post-translationally acetylates internal lysine residues of proteins. ARD1/NAA10 is the only enzyme with both NAT and KAT activities. However, recent studies on the role of human ARD1/NAA10 (hARD1/NAA10) in lysine acetylation are contradictory, as crystal structure and in vitro acetylation assay results revealed the lack of KAT activity. Thus, the role of hARD1/NAA10 in lysine acetylation is still debating. Here, we found a clue that possibly explains these complicated and controversial results on KAT activity of hARD1/NAA10. Recombinant hARD1/NAA10 exhibited KAT activity, which disappeared soon in vitro. Size-exclusion analysis revealed that most recombinant hARD1/NAA10 formed oligomers over time, resulting in the loss of KAT activity. While oligomeric recombinant hARD1/NAA10 lost its ability for lysine acetylation, its monomeric form clearly exhibited lysine acetylation activity in vitro. We also characterized the KAT activity of hARD1/NAA10 that was influenced by several experimental conditions, including concentration of reactants and reaction time. Taken together, our study proves that recombinant hARD1/NAA10 exhibits KAT activity in vitro but only under accurate conditions, including reactant concentrations and reaction duration.
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spelling pubmed-70368452020-03-11 Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10 Vo, Tam Thuy Lu Park, Ji-Hyeon Lee, Eun Ji Nguyen, Yen Thi Kim Han, Byung Woo Nguyen, Hien Thi Thu Mun, Kyo Cheol Ha, Eunyoung Kwon, Taeg Kyu Kim, Kyu-Won Jeong, Chul-Ho Seo, Ji Hae Molecules Article Arrest defective 1 (ARD1), also known as N(alpha)-acetyltransferase 10 (NAA10) was originally identified as an N-terminal acetyltransferase (NAT) that catalyzes the acetylation of N-termini of newly synthesized peptides. After that, mammalian ARD1/NAA10 expanded its’ role to lysine acetyltransferase (KAT) that post-translationally acetylates internal lysine residues of proteins. ARD1/NAA10 is the only enzyme with both NAT and KAT activities. However, recent studies on the role of human ARD1/NAA10 (hARD1/NAA10) in lysine acetylation are contradictory, as crystal structure and in vitro acetylation assay results revealed the lack of KAT activity. Thus, the role of hARD1/NAA10 in lysine acetylation is still debating. Here, we found a clue that possibly explains these complicated and controversial results on KAT activity of hARD1/NAA10. Recombinant hARD1/NAA10 exhibited KAT activity, which disappeared soon in vitro. Size-exclusion analysis revealed that most recombinant hARD1/NAA10 formed oligomers over time, resulting in the loss of KAT activity. While oligomeric recombinant hARD1/NAA10 lost its ability for lysine acetylation, its monomeric form clearly exhibited lysine acetylation activity in vitro. We also characterized the KAT activity of hARD1/NAA10 that was influenced by several experimental conditions, including concentration of reactants and reaction time. Taken together, our study proves that recombinant hARD1/NAA10 exhibits KAT activity in vitro but only under accurate conditions, including reactant concentrations and reaction duration. MDPI 2020-01-29 /pmc/articles/PMC7036845/ /pubmed/32013195 http://dx.doi.org/10.3390/molecules25030588 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Vo, Tam Thuy Lu
Park, Ji-Hyeon
Lee, Eun Ji
Nguyen, Yen Thi Kim
Han, Byung Woo
Nguyen, Hien Thi Thu
Mun, Kyo Cheol
Ha, Eunyoung
Kwon, Taeg Kyu
Kim, Kyu-Won
Jeong, Chul-Ho
Seo, Ji Hae
Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10
title Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10
title_full Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10
title_fullStr Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10
title_full_unstemmed Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10
title_short Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10
title_sort characterization of lysine acetyltransferase activity of recombinant human ard1/naa10
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7036845/
https://www.ncbi.nlm.nih.gov/pubmed/32013195
http://dx.doi.org/10.3390/molecules25030588
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