Structural basis of FpGalNase and its combination with FpGalNAcDeAc for efficient A-to-O blood group conversion
Abstract Transfusion safety and blood typing continue to present significant challenges in clinical practice, including risks of incorrect blood transfusions and blood shortages. One promising solution is the enzymatic conversion of all red blood cell (RBC) types into universal O-type RBCs. However,...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-01-01
|
| Series: | Experimental Hematology & Oncology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40164-025-00599-7 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832585976575164416 |
|---|---|
| author | Meiling Zhou Kaishan Luo Chao Su Yan Sun Zuyan Huang Shuo Ma Xun Gao Jiwei Wang Chen Zhang Pengcheng Han Guoqiu Wu |
| author_facet | Meiling Zhou Kaishan Luo Chao Su Yan Sun Zuyan Huang Shuo Ma Xun Gao Jiwei Wang Chen Zhang Pengcheng Han Guoqiu Wu |
| author_sort | Meiling Zhou |
| collection | DOAJ |
| description | Abstract Transfusion safety and blood typing continue to present significant challenges in clinical practice, including risks of incorrect blood transfusions and blood shortages. One promising solution is the enzymatic conversion of all red blood cell (RBC) types into universal O-type RBCs. However, the major obstacle to this strategy is the relatively low catalytic efficiency of the enzymes involved. In this study, we investigated two enzymes from Flavonifractor plautii, N-acetylgalactosamine deacetylase (FpGalNAcDeAc) and galactosaminidase (FpGalNase), which demonstrate synergistic activity in efficiently converting A-type RBCs to O-type. We optimized treatment conditions, achieving over 99% conversion in just five minutes using phosphate buffer saline and a 16 nM enzyme concentration. Additionally, we engineered two fusion proteins, FpGalNAcDeAc-FpGalNase and FpGalNase-FpGalNAcDeAc, which showed a 28-fold increase in catalytic efficiency compared to the enzyme mixture. Using cryo-electron microscopy, we resolved the full-length structure of FpGalNase, identifying critical active site residues involved in its catalytic mechanism. This study provides essential structural and biochemical insights for clinical applications in blood group conversion, offering a promising approach for producing universal O-type RBCs. |
| format | Article |
| id | doaj-art-2fdedcd4ab0f43df93dadd0c91c827ef |
| institution | Kabale University |
| issn | 2162-3619 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Experimental Hematology & Oncology |
| spelling | doaj-art-2fdedcd4ab0f43df93dadd0c91c827ef2025-01-26T12:18:59ZengBMCExperimental Hematology & Oncology2162-36192025-01-011411410.1186/s40164-025-00599-7Structural basis of FpGalNase and its combination with FpGalNAcDeAc for efficient A-to-O blood group conversionMeiling Zhou0Kaishan Luo1Chao Su2Yan Sun3Zuyan Huang4Shuo Ma5Xun Gao6Jiwei Wang7Chen Zhang8Pengcheng Han9Guoqiu Wu10Jiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityJiangsu Provincial Key Laboratory of Critical Care Medicine, Advanced Institute for Life and Health, Center of Clinical Laboratory Medicine, Department of Pharmacy, School of Medicine, Zhongda Hospital, Southeast UniversityAbstract Transfusion safety and blood typing continue to present significant challenges in clinical practice, including risks of incorrect blood transfusions and blood shortages. One promising solution is the enzymatic conversion of all red blood cell (RBC) types into universal O-type RBCs. However, the major obstacle to this strategy is the relatively low catalytic efficiency of the enzymes involved. In this study, we investigated two enzymes from Flavonifractor plautii, N-acetylgalactosamine deacetylase (FpGalNAcDeAc) and galactosaminidase (FpGalNase), which demonstrate synergistic activity in efficiently converting A-type RBCs to O-type. We optimized treatment conditions, achieving over 99% conversion in just five minutes using phosphate buffer saline and a 16 nM enzyme concentration. Additionally, we engineered two fusion proteins, FpGalNAcDeAc-FpGalNase and FpGalNase-FpGalNAcDeAc, which showed a 28-fold increase in catalytic efficiency compared to the enzyme mixture. Using cryo-electron microscopy, we resolved the full-length structure of FpGalNase, identifying critical active site residues involved in its catalytic mechanism. This study provides essential structural and biochemical insights for clinical applications in blood group conversion, offering a promising approach for producing universal O-type RBCs.https://doi.org/10.1186/s40164-025-00599-7Blood transfusionFpGalNAcDeAcFpGalNaseGalactosaminidaseN-acetylgalactosamine deacetylasecryo-EM structure |
| spellingShingle | Meiling Zhou Kaishan Luo Chao Su Yan Sun Zuyan Huang Shuo Ma Xun Gao Jiwei Wang Chen Zhang Pengcheng Han Guoqiu Wu Structural basis of FpGalNase and its combination with FpGalNAcDeAc for efficient A-to-O blood group conversion Experimental Hematology & Oncology Blood transfusion FpGalNAcDeAc FpGalNase Galactosaminidase N-acetylgalactosamine deacetylase cryo-EM structure |
| title | Structural basis of FpGalNase and its combination with FpGalNAcDeAc for efficient A-to-O blood group conversion |
| title_full | Structural basis of FpGalNase and its combination with FpGalNAcDeAc for efficient A-to-O blood group conversion |
| title_fullStr | Structural basis of FpGalNase and its combination with FpGalNAcDeAc for efficient A-to-O blood group conversion |
| title_full_unstemmed | Structural basis of FpGalNase and its combination with FpGalNAcDeAc for efficient A-to-O blood group conversion |
| title_short | Structural basis of FpGalNase and its combination with FpGalNAcDeAc for efficient A-to-O blood group conversion |
| title_sort | structural basis of fpgalnase and its combination with fpgalnacdeac for efficient a to o blood group conversion |
| topic | Blood transfusion FpGalNAcDeAc FpGalNase Galactosaminidase N-acetylgalactosamine deacetylase cryo-EM structure |
| url | https://doi.org/10.1186/s40164-025-00599-7 |
| work_keys_str_mv | AT meilingzhou structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT kaishanluo structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT chaosu structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT yansun structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT zuyanhuang structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT shuoma structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT xungao structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT jiweiwang structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT chenzhang structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT pengchenghan structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion AT guoqiuwu structuralbasisoffpgalnaseanditscombinationwithfpgalnacdeacforefficientatoobloodgroupconversion |