Enzyme Functional Annotation: from new microbial metabolism to mammalian antiviral defense mechanisms
The vast majority of proteins have not been subjected to experimental functional annotation, and a large proportion are misannotated. We describe an integrated pipeline that supports high-throughput cloning, expression, purification, structure determination and functional annotation. The utility of this pipeline is described for the discovery of new microbial metabolism, new metabolites and new chemistry. We have recently recapitulated our entire pipeline under anaerobic conditions, which provides a one-of-a-kind structural and functional genomics resource for oxygen-sensitive proteins. This anaerobic resource has allowed for the discovery of a new mammalian anti-flavivirus host defense mechanism. Future community-wide efforts and large-scale team-based consortia will be required to adequately address the “annotation vacuum”.