2IP1

Crystal Structure Analysis of S. cerevisiae Tryptophanyl tRNA Synthetase

Summary for 2IP1

• Entry DOI: 10.2210/pdb2ip1/pdb
• Related: 1O5T
• Descriptor: Tryptophanyl-tRNA synthetase, TETRAETHYLENE GLYCOL (3 entities in total)
• Functional Keywords: rossmann fold, structural genomics, psi-2, protein structure initiative, center for high-throughput structural biology, chtsb, ligase
• Biological source: Saccharomyces cerevisiae (baker's yeast)
• Cellular location: Cytoplasm: Q12109
• Total number of polymer chains: 1
• Total formula weight: 50196.13

Authors

Malkowski, M.G.,Center for High-Throughput Structural Biology (CHTSB) (deposition date: 2006-10-11, release date: 2007-06-26, Last modification date: 2024-02-21)

Primary citation

Malkowski, M.G.,Quartley, E.,Friedman, A.E.,Babulski, J.,Kon, Y.,Wolfley, J.,Said, M.,Luft, J.R.,Phizicky, E.M.,DeTitta, G.T.,Grayhack, E.J.
Blocking S-adenosylmethionine synthesis in yeast allows selenomethionine incorporation and multiwavelength anomalous dispersion phasing.
Proc.Natl.Acad.Sci.Usa, 104:6678-6683, 2007

PubMed Abstract: Saccharomyces cerevisiae is an ideal host from which to obtain high levels of posttranslationally modified eukaryotic proteins for x-ray crystallography. However, extensive replacement of methionine by selenomethionine for anomalous dispersion phasing has proven intractable in yeast. We report a general method to incorporate selenomethionine into proteins expressed in yeast based on manipulation of the appropriate metabolic pathways. sam1(-) sam2(-) mutants, in which the conversion of methionine to S-adenosylmethionine is blocked, exhibit reduced selenomethionine toxicity compared with wild-type yeast, increased production of protein during growth in selenomethionine, and efficient replacement of methionine by selenomethionine, based on quantitative mass spectrometry and x-ray crystallography. The structure of yeast tryptophanyl-tRNA synthetase was solved to 1.8 A by using multiwavelength anomalous dispersion phasing with protein that was expressed and purified from the sam1(-) sam2(-) strain grown in selenomethionine. Six of eight selenium residues were located in the structure.

PubMed: 17426150
DOI: 10.1073/pnas.0610337104

Experimental method

X-RAY DIFFRACTION (1.8 Å)