Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila β-N-acetylhexosaminidase Am2136
β-N-acetylhexosaminidases (EC3.2.1.52), which belong to the glycosyl hydrolase family GH20, are important enzymes for oligosaccharides modification. Numerous microbial β-N-acetylhexosaminidases have been investigated for applications in biology, biomedicine and biotechnology. Akkermansia muciniphila...
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Taylor & Francis Group
2022-12-01
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| Series: | Gut Microbes |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19490976.2022.2143221 |
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| author | Chang-Cheng Li Huan Yi Yan-Mei Wang Xin-Yue Tang Yi-Bo Zhu Ying-Jie Song Ning-Lin Zhao Qin Huang Xing-Yu Mou Gui-Hua Luo Tong-Gen Liu Gang-Long Yang Yu-Jiao Zeng Li-Jie Wang Hong Tang Gang Fan Rui Bao |
| author_facet | Chang-Cheng Li Huan Yi Yan-Mei Wang Xin-Yue Tang Yi-Bo Zhu Ying-Jie Song Ning-Lin Zhao Qin Huang Xing-Yu Mou Gui-Hua Luo Tong-Gen Liu Gang-Long Yang Yu-Jiao Zeng Li-Jie Wang Hong Tang Gang Fan Rui Bao |
| author_sort | Chang-Cheng Li |
| collection | DOAJ |
| description | β-N-acetylhexosaminidases (EC3.2.1.52), which belong to the glycosyl hydrolase family GH20, are important enzymes for oligosaccharides modification. Numerous microbial β-N-acetylhexosaminidases have been investigated for applications in biology, biomedicine and biotechnology. Akkermansia muciniphila is an anaerobic intestinal commensal bacterium which possesses specific β-N-acetylhexosaminidases for gut mucosal layer colonization and mucin degradation. In this study, we assessed the in vitro mucin glycan cleavage activity of the A. muciniphila β-N-acetylhexosaminidase Am2136 and demonstrated its ability that hydrolyzing the β-linkages joining N-acetylglucosamine to a wide variety of aglycone residues, which indicated that Am2136 may be a generalist β-N-acetylhexosaminidase. Structural and enzyme activity assay experiments allowed us to probe the essential function of the inter-domain interactions in β23-β33. Importantly, we revealed that the hydrolysis activity of Am2136 was enhanced by nucleotides. We further speculated that this activation mechanism might be associated with the conformational motions between domain III and IV. To our knowledge, this is the first report of nucleotide effector regulated β-N-acetylhexosaminidase, to reveal its novel biological functions. These findings contribute to understanding the distinct properties within the GH20 family and lay a certain foundation to develop controllable glycan hydrolyzing catalysts.Abbreviations: OD600 - optical cell densities at 600 nm; LB - Luria–Bertani; IPTG - isopropyl β-D-1-thiogalactopyranoside; PMSF - phenylmethanesulfonyl fluoride; rmsd - root mean square deviation; GlcNAc - N-acetyl-β-D-glucosamine; GalNAc - N-acetyl-β-D-galactosamine; Gal - galactose |
| format | Article |
| id | doaj-art-1760a3acb9b44aa8aeeb3cb848ff16d7 |
| institution | DOAJ |
| issn | 1949-0976 1949-0984 |
| language | English |
| publishDate | 2022-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Gut Microbes |
| spelling | doaj-art-1760a3acb9b44aa8aeeb3cb848ff16d72025-08-20T03:05:25ZengTaylor & Francis GroupGut Microbes1949-09761949-09842022-12-0114110.1080/19490976.2022.2143221Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila β-N-acetylhexosaminidase Am2136Chang-Cheng Li0Huan Yi1Yan-Mei Wang2Xin-Yue Tang3Yi-Bo Zhu4Ying-Jie Song5Ning-Lin Zhao6Qin Huang7Xing-Yu Mou8Gui-Hua Luo9Tong-Gen Liu10Gang-Long Yang11Yu-Jiao Zeng12Li-Jie Wang13Hong Tang14Gang Fan15Rui Bao16Division of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaState Key Laboratory of Southwestern Chinese Medicine Resources, College of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, ChinaInstitute of traditional Chinese medicine, Sichuan College of traditional Chinese Medicine (Sichuan Second Hospital of TCM), Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaSchool of Biotechnology, Jiangnan University, Chengdu, ChinaState Key Laboratory of Southwestern Chinese Medicine Resources, College of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, ChinaState Key Laboratory of Southwestern Chinese Medicine Resources, College of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, ChinaState Key Laboratory of Southwestern Chinese Medicine Resources, College of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, ChinaDivision of Infectious Diseases, State Key Laboratory of Biotherapy and Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, Chinaβ-N-acetylhexosaminidases (EC3.2.1.52), which belong to the glycosyl hydrolase family GH20, are important enzymes for oligosaccharides modification. Numerous microbial β-N-acetylhexosaminidases have been investigated for applications in biology, biomedicine and biotechnology. Akkermansia muciniphila is an anaerobic intestinal commensal bacterium which possesses specific β-N-acetylhexosaminidases for gut mucosal layer colonization and mucin degradation. In this study, we assessed the in vitro mucin glycan cleavage activity of the A. muciniphila β-N-acetylhexosaminidase Am2136 and demonstrated its ability that hydrolyzing the β-linkages joining N-acetylglucosamine to a wide variety of aglycone residues, which indicated that Am2136 may be a generalist β-N-acetylhexosaminidase. Structural and enzyme activity assay experiments allowed us to probe the essential function of the inter-domain interactions in β23-β33. Importantly, we revealed that the hydrolysis activity of Am2136 was enhanced by nucleotides. We further speculated that this activation mechanism might be associated with the conformational motions between domain III and IV. To our knowledge, this is the first report of nucleotide effector regulated β-N-acetylhexosaminidase, to reveal its novel biological functions. These findings contribute to understanding the distinct properties within the GH20 family and lay a certain foundation to develop controllable glycan hydrolyzing catalysts.Abbreviations: OD600 - optical cell densities at 600 nm; LB - Luria–Bertani; IPTG - isopropyl β-D-1-thiogalactopyranoside; PMSF - phenylmethanesulfonyl fluoride; rmsd - root mean square deviation; GlcNAc - N-acetyl-β-D-glucosamine; GalNAc - N-acetyl-β-D-galactosamine; Gal - galactosehttps://www.tandfonline.com/doi/10.1080/19490976.2022.2143221Akkermansia muciniphilaglycosidasemucinprotein structure |
| spellingShingle | Chang-Cheng Li Huan Yi Yan-Mei Wang Xin-Yue Tang Yi-Bo Zhu Ying-Jie Song Ning-Lin Zhao Qin Huang Xing-Yu Mou Gui-Hua Luo Tong-Gen Liu Gang-Long Yang Yu-Jiao Zeng Li-Jie Wang Hong Tang Gang Fan Rui Bao Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila β-N-acetylhexosaminidase Am2136 Gut Microbes Akkermansia muciniphila glycosidase mucin protein structure |
| title | Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila β-N-acetylhexosaminidase Am2136 |
| title_full | Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila β-N-acetylhexosaminidase Am2136 |
| title_fullStr | Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila β-N-acetylhexosaminidase Am2136 |
| title_full_unstemmed | Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila β-N-acetylhexosaminidase Am2136 |
| title_short | Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila β-N-acetylhexosaminidase Am2136 |
| title_sort | nucleotide binding as an allosteric regulatory mechanism for akkermansia muciniphila β n acetylhexosaminidase am2136 |
| topic | Akkermansia muciniphila glycosidase mucin protein structure |
| url | https://www.tandfonline.com/doi/10.1080/19490976.2022.2143221 |
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