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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...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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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. |
format | Online Article Text |
id | pubmed-7036845 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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