Bottlenecks and roadblocks in high-throughput XAS for structural genomics.
Structural and functional characterization of the entire protein complement (the proteome) of an organism can provide an infrastructure upon which questions about biological pathways and systems biology can be framed. The technology necessary to perform this proteome-level structural and functional characterization is under development in numerous structural genomics and functional genomics initiatives. Given the ubiquity of metal active sites in a proteome, it seems appropriate to ask whether comprehensive local structural characterization of metal sites within a proteome (metalloproteomics) is either a valid or obtainable goal. With a proteome-wide knowledge of the active-site structures of all metalloproteins, one could start to ask how metal insertion, cluster assembly and metalloprotein expression are affected by growth conditions or developmental status etc. High-throughput X-ray absorption spectroscopy (HTXAS) is being developed as a technology for investigating the metalloproteome. In creating a pipeline from genome to metalloproteome, several bottlenecks to high-throughput determination of metal-site structures must be overcome. For example, automation of arraying small samples for XAS examination must be invented, automation of rapid data collection of multiple low-volume low-concentration samples must be developed, automation of data reduction and analysis must be perfected. Discussed here are the promises and the pitfalls of HTXAS development, including the results of initial feasibility experiments.
Scott
Robert A
RA
Department of Chemistry, University of Georgia, Athens, GA 30602, USA. scott@chem.uga.edu
Shokes
Jacob E
JE
Cosper
Nathaniel J
NJ
Jenney
Francis E
FE
Adams
Michael W W
MW
D001331:Automation; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics*; D003001:Cloning, Molecular; D005240:Feasibility Studies; D016680:Genome, Bacterial; D008667:Metalloproteins / Q000737:chemistry* / Q000235:genetics*; D011487:Protein Conformation; D040901:Proteomics; D013057:Spectrum Analysis / Q000379:methods*; D014965:X-Rays
D001426:Bacterial Proteins; D008667:Metalloproteins