Characterization of metalloproteins by high-throughput X-ray absorption spectroscopy.

摘要: High-throughput X-ray absorption spectroscopy was used to measure transition metal content based on quantitative detection of fluorescence signals for 3879 purified proteins from several hundred different protein families generated by the New York SGX Research Center Structural Genomics. Approximately 9% analyzed showed presence atoms (Zn, Cu, Ni, Co, Fe, or Mn) in stoichiometric amounts. The method is highly automated and reliable comparison results crystal structure data derived same set. To leverage experimental metalloprotein annotations, we a sequence-based de novo prediction method, MetalDetector, identify Cys His residues that bind metals redundancy reduced subset 2411 sequences sharing <70% sequence identity having at least one Cys. As HT-XAS identifies type binding, while bioinformatics analysis metal- binding residues, were combined putative metal-binding sites their associated families. We explored combination this with homology models generate detailed representative proteins. Finally, extended fine two Zn metalloproteins validate predicted site structures. This approaches provides comprehensive active genome scale as class, revealing new insights into function.