High throughput technique in structural bioinformatics

Abstract

High Throughput techniques are necessary for solvin g the three dimensional structures of macromolecule s because the macromolecular sequences available exceed far i n number than the available three-dimensional struc ture. Experimental phasing using anomalous data is essential to solve non-homologous protein structures. Su lfur atom is commonly present in protein structures in the form of cysteine and methionine residues. With recent pr ogress in data collection, Sulfur ‐ SAD phasing methods are extensively used for phasing macromolecular structures. S -SAD phasing has been tested on experimental Cr K α data set, 2.4 A, of an enzyme glucose isomerase of molecular weight 44 kDa. The combination of PHENIX and ARP/wARP built 379 residues out of 388 residues using 11 sulfurs located initially from the SAD data. Truncate the d ata to 3 A resolution for testing the power of S-SA D. The above combination built 283 residues out of 388 residues using 11 sulfurs. Successive model could be built u sing OASIS which built 375 residues out of 388 residues. Minim al manual model building was required at this stage and the structure determination was completed using REFMAC5.