Rational design of a structural and functional nitric oxide reductase
Open Access
- 25 November 2009
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature
- Vol. 462 (7276), 1079-1082
- https://doi.org/10.1038/nature08620
Abstract
Considerable progress has been made in the design of proteins that mimic native proteins both structurally and functionally. Metalloproteins present more of a challenge owing to their complexity, but Yi Lu and colleagues now report the successful design of a structural and functional model of the metalloprotein nitric oxide reductase (NOR). An X-ray crystal structure of the designed protein confirms that it contains a haem/non-haem FeB centre very similar to that in the native protein, and also exhibits NOR activity. Despite the progress that has been made in designing proteins that mimic native proteins structurally, it is difficult to design functional proteins and particularly challenging to design metalloproteins that reproduce both the structure and function of native metalloenzymes. Here, the successful, rational design of a structural and functional model of a metalloprotein — nitric oxide reductase — is achieved. Protein design provides a rigorous test of our knowledge about proteins and allows the creation of novel enzymes for biotechnological applications. Whereas progress has been made in designing proteins that mimic native proteins structurally1,2,3, it is more difficult to design functional proteins4,5,6,7,8. In comparison to recent successes in designing non-metalloproteins4,6,7,9,10, it is even more challenging to rationally design metalloproteins that reproduce both the structure and function of native metalloenzymes5,8,11,12,13,14,15,16,17,18,19,20. This is because protein metal-binding sites are much more varied than non-metal-containing sites, in terms of different metal ion oxidation states, preferred geometry and metal ion ligand donor sets. Because of their variability, it has been difficult to predict metal-binding site properties in silico, as many of the parameters, such as force fields, are ill-defined. Therefore, the successful design of a structural and functional metalloprotein would greatly advance the field of protein design and our understanding of enzymes. Here we report a successful, rational design of a structural and functional model of a metalloprotein, nitric oxide reductase (NOR), by introducing three histidines and one glutamate, predicted as ligands in the active site of NOR, into the distal pocket of myoglobin. A crystal structure of the designed protein confirms that the minimized computer model contains a haem/non-haem FeB centre that is remarkably similar to that in the crystal structure. This designed protein also exhibits NO reduction activity, and so models both the structure and function of NOR, offering insight that the active site glutamate is required for both iron binding and activity. These results show that structural and functional metalloproteins can be rationally designed in silico.This publication has 43 references indexed in Scilit:
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