Three-dimensional Structure of YaaE from Bacillus subtilis, a Glutaminase Implicated in Pyridoxal-5′-phosphate Biosynthesis*

The structure of YaaE from Bacillus subtilis was determined at 2.5-Å resolution. YaaE is a member of the triad glutamine aminotransferase family and functions in a recently identified alternate pathway for the biosynthesis of vitamin B6. Proposed active residues include conserved Cys-79, His-170, and Glu-172. YaaE shows similarity to HisH, a glutaminase involved in histidine biosynthesis. YaaD associates with YaaE. A homology model of this protein was constructed. YaaD is predicted to be a (β/α)8 barrel on the basis of sequence comparisons. The predicted active site includes highly conserved residues 211–216 and 233–235. Finally, a homology model of a putative YaaD-YaaE complex was prepared using the structure of HisH-F as a model. This model predicts that the ammonia molecule generated by YaaE is channeled through the center of the YaaD barrel to the putative YaaD active site.

[1]  W. L. Jorgensen,et al.  Development and Testing of the OPLS All-Atom Force Field on Conformational Energetics and Properties of Organic Liquids , 1996 .

[2]  Janet L. Smith,et al.  The crystal structure of GMP synthetase reveals a novel catalytic triad and is a structural paradigm for two enzyme families , 1996, Nature Structural Biology.

[3]  C. Drewke,et al.  Biosynthesis of vitamin B6 and structurally related derivatives. , 2001, Vitamins and hormones.

[4]  P. Kraulis A program to produce both detailed and schematic plots of protein structures , 1991 .

[5]  J. Zeidler,et al.  Biosynthesis of Vitamin B(6) in yeast. Incorporation pattern of trioses. , 2003, The Journal of organic chemistry.

[6]  W. C. Still,et al.  The GB/SA Continuum Model for Solvation. A Fast Analytical Method for the Calculation of Approximate Born Radii , 1997 .

[7]  M. Werner-Washburne,et al.  The Highly Conserved, Coregulated SNOand SNZ Gene Families in Saccharomyces cerevisiaeRespond to Nutrient Limitation , 1998, Journal of bacteriology.

[8]  T. N. Bhat,et al.  The Protein Data Bank , 2000, Nucleic Acids Res..

[9]  J. Bernhardt,et al.  First steps from a two‐dimensional protein index towards a response‐regulation map for Bacillus subtilis , 1997, Electrophoresis.

[10]  M. Werner-Washburne,et al.  A stationary-phase gene in Saccharomyces cerevisiae is a member of a novel, highly conserved gene family , 1996, Journal of bacteriology.

[11]  M. Werner-Washburne,et al.  Analysis of the pdx-1 (snz-1/sno-1) region of the Neurospora crassa genome: correlation of pyridoxine-requiring phenotypes with mutations in two structural genes. , 2001, Genetics.

[12]  S. Osmani,et al.  The Extremely Conserved pyroA Gene ofAspergillus nidulans Is Required for Pyridoxine Synthesis and Is Required Indirectly for Resistance to Photosensitizers* , 1999, The Journal of Biological Chemistry.

[13]  E A Merritt,et al.  Raster3D: photorealistic molecular graphics. , 1997, Methods in enzymology.

[14]  M. Sternberg,et al.  Enhanced genome annotation using structural profiles in the program 3D-PSSM. , 2000, Journal of molecular biology.

[15]  M. Werner-Washburne,et al.  Protein synthesis in long-term stationary-phase cultures of Saccharomyces cerevisiae , 1994, Journal of bacteriology.

[16]  T. Blundell,et al.  Comparative protein modelling by satisfaction of spatial restraints. , 1993, Journal of molecular biology.

[17]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[18]  Sridar V. Chittur,et al.  Crystal Structure of Imidazole Glycerol Phosphate Synthase: A Tunnel through a (β/α)8 Barrel Joins Two Active Sites , 2001 .

[19]  Tohoru Katsuragi,et al.  yaaD and yaaE are involved in vitamin B6 biosynthesis in Bacillus subtilis. , 2002, Journal of bioscience and bioengineering.

[20]  H M Holden,et al.  Carbamoyl phosphate synthetase: caught in the act of glutamine hydrolysis. , 1998, Biochemistry.

[21]  K. Chung,et al.  Functional characterization of SOR1, a gene required for resistance to photosensitizing toxins in the fungus Cercospora nicotianae , 1999, Current Genetics.

[22]  J. Zeidler,et al.  2'-hydroxypyridoxol, a biosynthetic precursor of vitamins B(6) and B(1) in yeast. , 2002, Journal of the American Chemical Society.

[23]  M. Daub,et al.  Isolation of PDX2, a Second Novel Gene in the Pyridoxine Biosynthesis Pathway of Eukaryotes, Archaebacteria, and a Subset of Eubacteria , 2001, Journal of bacteriology.

[24]  C. Yanofsky,et al.  The structures of anthranilate synthase of Serratia marcescens crystallized in the presence of (i) its substrates, chorismate and glutamine, and a product, glutamate, and (ii) its end-product inhibitor, l-tryptophan , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[26]  H. Zalkin,et al.  Replacement by site-directed mutagenesis indicates a role for histidine 170 in the glutamine amide transfer function of anthranilate synthase. , 1985, The Journal of biological chemistry.

[27]  A. Sali,et al.  Comparative Protein Structure Prediction , 2002 .

[28]  K. Yamada,et al.  Origin of the nitrogen atom of pyridoxine in Saccharomyces cerevisiae. , 1995, Biochimica et biophysica acta.

[29]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[30]  P E Bourne,et al.  Protein structure alignment by incremental combinatorial extension (CE) of the optimal path. , 1998, Protein engineering.

[31]  Z. Otwinowski,et al.  [20] Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[32]  G. Mittenhuber,et al.  Phylogenetic analyses and comparative genomics of vitamin B6 (pyridoxine) and pyridoxal phosphate biosynthesis pathways. , 2001, Journal of molecular microbiology and biotechnology.

[33]  Chris Sander,et al.  Touring protein fold space with Dali/FSSP , 1998, Nucleic Acids Res..

[34]  Matthias Wilmanns,et al.  Structural evidence for ammonia tunneling across the (beta alpha)(8) barrel of the imidazole glycerol phosphate synthase bienzyme complex. , 2002, Structure.

[35]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.