(A) Workflow for building a comprehensive in silico erythrocyte metabolic network. The three major data types required are: the human genome sequence, high-throughput data (specifically, proteomics for an enucleated cell), and primary literature. The global human metabolic network, Recon 1, was constructed from the human genome sequence and annotation. To build the erythrocyte network, iAB-RBC-283, proteomics was used to remove non-erythrocyte related open reading frames (ORFs) or genes. Detailed curation utilizing protein, metabolite, and transport experimental literature was needed to build a high-quality metabolic reconstruction. (B) Without network reconstruction and rigorous curation, the experimentally generated proteomic data is raw and difficult to interpret. The process detailed in panel A provides a means to build a meaningful knowledge-base of available high-throughput data that can then be probed and tested.