期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 288, 期 40, 页码 29081-29089出版社
ELSEVIER
DOI: 10.1074/jbc.M113.497412
关键词
Cellulase; Computational Biology; Enzyme Inactivation; Enzyme Kinetics; Macromolecular Crowding; Molecular Modeling; Systems Biology; Interfacial Biocatalysis; Spatiotemporal Modeling
资金
- Office of Science of the United States Department of Energy [DE-AC02-05CH11231]
- National Research Foundation of Korea [2013M1A8A1038187, 2012M3C1A6035363] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Interprotein and enzyme-substrate couplings in interfacial biocatalysis induce spatial correlations beyond the capabilities of classical mass-action principles in modeling reaction kinetics. To understand the impact of spatial constraints on enzyme kinetics, we developed a computational scheme to simulate the reaction network of enzymes with the structures of individual proteins and substrate molecules explicitly resolved in the three-dimensional space. This methodology was applied to elucidate the rate-limiting mechanisms of crystalline cellulose decomposition by cellobiohydrolases. We illustrate that the primary bottlenecks are slow complexation of glucan chains into the enzyme active site and excessive enzyme jamming along the crowded substrate. Jamming could be alleviated by increasing the decomplexation rate constant but at the expense of reduced processivity. We demonstrate that enhancing the apparent reaction rate required a subtle balance between accelerating the complexation driving force and simultaneously avoiding enzyme jamming. Via a spatiotemporal systems analysis, we developed a unified mechanistic framework that delineates the experimental conditions under which different sets of rate-limiting behaviors emerge. We found that optimization of the complexation-exchange kinetics is critical for overcoming the barriers imposed by interfacial confinement and accelerating the apparent rate of enzymatic cellulose decomposition.
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