3sds | pdb_00003sds

Crystal structure of a mitochondrial ornithine carbamoyltransferase from Coccidioides immitis

spinqualitylabelsall models 3sds, resolution 2.80Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 3sds is a 3 chain structure with sequence from Coccidioides immitis. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Ligands: CL Gene: CIMG_04084 (Coccidioides immitis) Activity: Ornithine carbamoyltransferase, with EC number 2.1.3.3 Resources: FirstGlance, OCA, RCSB, PDBsum Publication Abstract from PubMed Despite recent advances, the expression of heterologous proteins in Escherichia coli for crystallization remains a nontrivial challenge. The present study investigates the efficacy of maltose-binding protein (MBP) fusion as a general strategy for rescuing the expression of target proteins. From a group of sequence-verified clones with undetectable levels of protein expression in an E. coli T7 expression system, 95 clones representing 16 phylogenetically diverse organisms were selected for recloning into a chimeric expression vector with an N-terminal histidine-tagged MBP. PCR-amplified inserts were annealed into an identical ligation-independent cloning region in an MBP-fusion vector and were analyzed for expression and solubility by high-throughput nickel-affinity binding. This approach yielded detectable expression of 72% of the clones; soluble expression was visible in 62%. However, the solubility of most proteins was marginal to poor upon cleavage of the MBP tag. This study offers large-scale evidence that MBP can improve the soluble expression of previously non-expressing proteins from a variety of eukaryotic and prokaryotic organisms. While the behavior of the cleaved proteins was disappointing, further refinements in MBP tagging may permit the more widespread use of MBP-fusion proteins in crystallographic studies. Expression of proteins in Escherichia coli as fusions with maltose-binding protein to rescue non-expressed targets in a high-throughput protein-expression and purification pipeline.,Hewitt SN, Choi R, Kelley A, Crowther GJ, Napuli AJ, Van Voorhis WC Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Sep 1;67(Pt, 9):1006-9. Epub 2011 Aug 13. PMID:21904041 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See Also Ornithine carbamoyltransferase

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