Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin
Abstract
:1. Introduction
2. Results
2.1. MitE Catalyzed the CoA Ligation of AHBA by the Genetic and Biochemcal Analysis
2.2. MmcB-Tethering AHBA Triggered the Biosynthesis of Mitomycin C
2.3. The Completed MitB Catalyzed the Glycosylation of ACP-Bound AHBA
2.4. The Protein–Protein Interactions Occurred between MmcB and MitB
2.5. MmcB Interacted with MitF during Mitomycin C Assembly
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains, Plasmids, and Culture Conditions
4.2. Gene Deletion and Complementation
4.2.1. Gene MitE In-Frame Deletion and Complementation
4.2.2. Gene MmcB In-Frame Deletion and Complementation
4.2.3. Gene MitB In-Frame Deletion and Complementation
4.2.4. Gene MitF In-Frame Deletion and Complementation
4.3. Production and Detection of Mitomycin C in Strain NRRL2564
4.4. Protein Expression and Purification
4.5. Enzyme Activity Analysis
4.6. Determination of the Protein Molecular Weight by Q-TOF Mass Spectrometry
4.7. Protein–Protein Interaction Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Leng, D.; Sheng, Y.; Wang, H.; Wei, J.; Ou, Y.; Deng, Z.; Bai, L.; Kang, Q. Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin. Molecules 2021, 26, 6791. https://doi.org/10.3390/molecules26226791
Leng D, Sheng Y, Wang H, Wei J, Ou Y, Deng Z, Bai L, Kang Q. Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin. Molecules. 2021; 26(22):6791. https://doi.org/10.3390/molecules26226791
Chicago/Turabian StyleLeng, Dongjin, Yong Sheng, Hengyu Wang, Jianhua Wei, Yixin Ou, Zixin Deng, Linquan Bai, and Qianjin Kang. 2021. "Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin" Molecules 26, no. 22: 6791. https://doi.org/10.3390/molecules26226791