Structure and function of Mycobacterium smegmatis 7-keto-8-aminopelargonic acid (KAPA) synthase |
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| Institution: | 1. State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China;2. National Laboratory of Biomacromolecules and CAS Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;3. Center for Tuberculosis Control of Guangdong Province, Guangdong 510630, China;4. The 4th People''s Hospital, Foshan, Guangdong 528000, China;1. School of Environment, Henan Normal University, Key Laboratory for Yellow River and Huai River Water Environmental and Pollution Control, Ministry of Education, Henan Key Laboratory for Environmental Pollution Control, Xinxiang, Henan 453007, PR China;2. Center for Nanochemistry (CNC), College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, PR China;1. Department of Chemistry, D.S.B Campus, Kumaun University, Nainital 260002, Uttarakhand;2. Department of Biotechnology, Sir J. C. Bose Technical Campus, Bhimtal, Kumaun University, Nainital 263136, Uttarakhand;1. Department of Chemical Engineering and Technology, Indian Institute of Technology (Banaras Hindu University), Varanasi 221005, India;2. Gasification and Liquefaction Research Group, CSIR-Central Institute of Mining and Fuel Research (DC), Dhanbad 828108, India;1. Key Laboratory of Marine Environmental Corrosion and Bio-fouling, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, China;2. University of Chinese Academy of Sciences, 19 (Jia) Yuquan Road, Beijing 100039, China;3. Laboratory of Clean Energy Chemistry and Materials, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, 18 Tianshui Middle Road, Lanzhou 730000, China;1. Key Laboratory of Green Catalysis of Higher Education Institutes of Sichuan, College of Chemistry and Environmental Engineering, Sichuan University of Science and Engineering, Zigong, 643000, PR China;2. Department of Chemistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, United States |
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| Abstract: | The biotin biosynthesis pathway is an attractive target for development of novel drugs against mycobacterial pathogens, however there are as yet no suitable inhibitors that target this pathway in mycobacteria. 7-Keto-8-aminopelargonic acid synthase (KAPA synthase, BioF) is the enzyme which catalyzes the first committed step of the biotin synthesis pathway, but both its structure and function in mycobacteria remain unresolved. Here we present the crystal structure of Mycobacterium smegmatis BioF (MsBioF). The structure reveals an incomplete dimer, and the active site organization is similar to, but distinct from Escherichia coli 8-amino-7-oxononanoate synthase (EcAONS), the E. coli homologue of BioF. To investigate the influence of structural characteristics on the function of MsBioF, we deleted bioF in M. smegmatis and confirmed that BioF is required for growth in the absence of exogenous biotin. Based on structural and mutagenesis studies, we confirmed that pyridoxal 5′-phosphate (PLP) binding site residues His129, Lys235 and His200 are essential for MsBioF activity in vivo and residue Glu171 plays an important, but not essential role in MsBioF activity. The N-terminus (residues 1–37) is also essential for MsBioF activity in vivo. The structure and function of MsBioF reported here provides further insights for developing new anti-tuberculosis inhibitors aimed at the biotin synthesis pathway. |
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| Keywords: | Biotin synthesis pathway 7-Keto-8-aminopelargonic acid (KAPA) synthase Crystal structure Active site |
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