Identifying protein domains by global analysis of soluble fragment data.

摘要: Abstract The production and analysis of individual structural domains is a common strategy for studying large or complex proteins, which may be experimentally intractable in their full-length form. However, identifying domain boundaries challenging if there little information concerning the protein target. One experimental procedure mapping to screen library random fragments solubility, since truncation will typically expose hydrophobic groups, leading poor fragment solubility. We have coupled solubility screening with global data develop an effective method within protein. A gene generated using mechanical shearing, by uracil doping uracil-specific enzymatic digest. split green fluorescent (GFP) assay used corresponding when expressed Escherichia coli . soluble are then analyzed two complementary approaches. Fragmentation “hotspots” indicate possible interdomain regions. Clustering algorithms group related fragments, concomitantly predict location. effectiveness this Domain Seeking demonstrated application well-characterized human p85α.