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 acetylt...
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| Format: | Article |
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MDPI AG
2020-01-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/3/588 |
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| author | Tam Thuy Lu Vo Ji-Hyeon Park Eun Ji Lee Yen Thi Kim Nguyen Byung Woo Han Hien Thi Thu Nguyen Kyo Cheol Mun Eunyoung Ha Taeg Kyu Kwon Kyu-Won Kim Chul-Ho Jeong Ji Hae Seo |
| author_facet | Tam Thuy Lu Vo Ji-Hyeon Park Eun Ji Lee Yen Thi Kim Nguyen Byung Woo Han Hien Thi Thu Nguyen Kyo Cheol Mun Eunyoung Ha Taeg Kyu Kwon Kyu-Won Kim Chul-Ho Jeong Ji Hae Seo |
| author_sort | Tam Thuy Lu Vo |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-21T17:17:57Z |
| format | Article |
| id | doaj.art-1d53217553d342b093883deb22f87d46 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-21T17:17:57Z |
| publishDate | 2020-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-1d53217553d342b093883deb22f87d462022-12-21T18:56:15ZengMDPI AGMolecules1420-30492020-01-0125358810.3390/molecules25030588molecules25030588Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10Tam Thuy Lu Vo0Ji-Hyeon Park1Eun Ji Lee2Yen Thi Kim Nguyen3Byung Woo Han4Hien Thi Thu Nguyen5Kyo Cheol Mun6Eunyoung Ha7Taeg Kyu Kwon8Kyu-Won Kim9Chul-Ho Jeong10Ji Hae Seo11Department of Biochemistry, Keimyung University School of Medicine, Daegu 42601, KoreaDepartments of Radiology and Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA 02129, USADepartments of Radiology and Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA 02129, USACollege of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul 08826, KoreaCollege of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul 08826, KoreaDepartment of Biochemistry, Keimyung University School of Medicine, Daegu 42601, KoreaDepartment of Biochemistry, Keimyung University School of Medicine, Daegu 42601, KoreaDepartment of Biochemistry, Keimyung University School of Medicine, Daegu 42601, KoreaDepartment of Immunology, Keimyung University School of Medicine, Daegu 42601, KoreaCollege of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul 08826, KoreaCollege of Pharmacy, Keimyung University, Daegu 42601, KoreaDepartment of Biochemistry, Keimyung University School of Medicine, Daegu 42601, KoreaArrest 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.https://www.mdpi.com/1420-3049/25/3/588arrest defective 1 (ard1)acetylationn(alpha)-acetyltransferase 10 (naa10)lysine acetyltransferase (kat)n-terminal acetyltransferase (nat) |
| spellingShingle | Tam Thuy Lu Vo Ji-Hyeon Park Eun Ji Lee Yen Thi Kim Nguyen Byung Woo Han Hien Thi Thu Nguyen Kyo Cheol Mun Eunyoung Ha Taeg Kyu Kwon Kyu-Won Kim Chul-Ho Jeong Ji Hae Seo Characterization of Lysine Acetyltransferase Activity of Recombinant Human ARD1/NAA10 Molecules arrest defective 1 (ard1) acetylation n(alpha)-acetyltransferase 10 (naa10) lysine acetyltransferase (kat) n-terminal acetyltransferase (nat) |
| 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 | arrest defective 1 (ard1) acetylation n(alpha)-acetyltransferase 10 (naa10) lysine acetyltransferase (kat) n-terminal acetyltransferase (nat) |
| url | https://www.mdpi.com/1420-3049/25/3/588 |
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