期刊
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
卷 144, 期 16, 页码 7146-7159出版社
AMER CHEMICAL SOC
DOI: 10.1021/jacs.1c12629
关键词
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资金
- Generalitat de Catalunya
- Spanish MINECO [PGC2018-102192-B-I00, RTI2018-101032-J100, RYC2020-029552-I]
- European Research Council (ERC) under the European Union [ERC-2015-StG-679001]
- Human Frontier Science Program (HFSP) [RGP0054/2020, BCV-2021-1-0015]
- National Research Foundation of Korea (NRF) under the Brain Pool Program [NRF-2021H1D3A2A02038434]
- ACS
- National Research Foundation of Korea [2021H1D3A2A02038434] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
This research investigates the allosteric activation mechanisms of IGPS using molecular dynamics simulations, enhanced sampling techniques, and dynamical networks. The simulations reveal how IGPS captures the substrate, forms a closed interface, and creates an oxyanion hole for efficient substrate hydrolysis under the influence of the allosteric effector. The study provides insights into the molecular details of allosteric activation and can guide engineering efforts of IGPS.
Deciphering the molecular mechanisms of enzymatic allosteric regulation requires the structural characterization of functional states and also their time evolution toward the formation of the allosterically activated ternary complex. The transient nature and usually slow millisecond time scale interconversion between these functional states hamper their experimental and computational characterization. Here, we combine extensive molecular dynamics simulations, enhanced sampling techniques, and dynamical networks to describe the allosteric activation of imidazole glycerol phosphate synthase (IGPS) from the substrate-free form to the active ternary complex. IGPS is a heterodimeric bienzyme complex whose HisH subunit is responsible for hydrolyzing glutamine and delivering ammonia for the cyclase activity in HisF. Despite significant advances in understanding the underlying allosteric mechanism, essential molecular details of the long-range millisecond allosteric activation of IGPS remain hidden. Without using a priori information of the active state, our simulations uncover how IGPS, with the allosteric effector bound in HisF, spontaneously captures glutamine in a catalytically inactive HisH conformation, subsequently attains a closed HisF:HisH interface, and finally forms the oxyanion hole in HisH for efficient glutamine hydrolysis. We show that the combined effector and substrate binding dramatically decreases the conformational barrier associated with oxyanion hole formation, in line with the experimentally observed 4500-fold activity increase in glutamine hydrolysis. The allosteric activation is controlled by correlated time-evolving dynamic networks connecting the effector and substrate binding sites. This computational strategy tailored to describe millisecond events can be used to rationalize the effect of mutations on the allosteric regulation and guide IGPS engineering efforts.
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