Atomic structure of thymidylate synthase: target for rational drug design.

The atomic structure of thymidylate synthase from Lactobacillus casei was determined at 3 angstrom resolution. The native enzyme is a dimer of identical subunits. The dimer interface is formed by an unusual association between five-stranded beta sheets present in each monomer. Comparison of known sequences with the Lactobacillus casei structure suggests that they all have a common core structure around which loops are inserted or deleted in different sequences. Residues from both subunits contribute to each active site. Two arginine side chains can contribute to binding phosphate on the substrate. The side chains of several conserved amino acids can account for other determinants of substrate binding.

[1]  L. Johnson,et al.  Structure of the gene for mouse thymidylate synthase. Locations of introns and multiple transcriptional start sites. , 1986, The Journal of biological chemistry.

[2]  L. Lebioda,et al.  Crystallization and crystallographic data for new forms of thymidylate synthase from Lactobacillus casei. , 1986, Journal of molecular biology.

[3]  D. Caput,et al.  Bifunctional thymidylate synthase-dihydrofolate reductase from Leishmania tropica: sequence homology with the corresponding monofunctional proteins. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[4]  R. Randall,et al.  The A+T-rich genome of Herpesvirus saimiri contains a highly conserved gene for thymidylate synthase. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[5]  S. Beverley,et al.  Primary structure of the gene encoding the bifunctional dihydrofolate reductase-thymidylate synthase of Leishmania major. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[6]  D. Santi,et al.  Limited proteolysis of the bifunctional thymidylate synthase-dihydrofolate reductase from Leishmania tropica. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[7]  O. Gotoh,et al.  Nucleic Acids Research Nucleotide sequence of a functional cDNA for human thymldylate synthase , 2004 .

[8]  R. Hamlin,et al.  [27] Multiwire area X-ray diffractometers , 1985 .

[9]  B. C. Wang.  Resolution of phase ambiguity in macromolecular crystallography. , 1985, Methods in enzymology.

[10]  N. Xuong,et al.  Software for a diffractometer with multiwire area detector. , 1985, Methods in enzymology.

[11]  T. Atkinson,et al.  Nucleotide sequence of the thymidylate synthetase gene (thyP3) from the Bacillus subtilis phage phi 3T. , 1985, Gene.

[12]  J S Richardson,et al.  Describing patterns of protein tertiary structure. , 1985, Methods in enzymology.

[13]  P. Danenberg,et al.  Spectroscopic studies of ternary complexes of thymidylate synthetase, deoxyribonucleotides, and folate analogs. , 1984, The Journal of biological chemistry.

[14]  M. Belfort,et al.  Intervening sequence in the thymidylate synthase gene of bacteriophage T4. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[15]  S. Beverley,et al.  A bifunctional thymidylate synthetase-dihydrofolate reductase in protozoa. , 1984, Molecular and biochemical parasitology.

[16]  R M Stroud,et al.  Amphipathic analysis and possible formation of the ion channel in an acetylcholine receptor. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[17]  M. Belfort,et al.  Primary structure of the Escherichia coli thyA gene and its thymidylate synthase product. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[18]  W. Rutter,et al.  Splice junctions: association with variation in protein structure. , 1983, Science.

[19]  D. Santi,et al.  Secondary alpha-hydrogen isotope effects on the interaction of 5-fluoro-2'-deoxyuridylate and 5,10-methylenetetrahydrofolate with thymidylate synthetase. , 1982, Biochemistry.

[20]  G. Maley,et al.  Studies on identifying the folylpolyglutamate binding sites of Lactobacillus casei thymidylate synthetase. , 1982, Archives of biochemistry and biophysics.

[21]  William R. Taylor,et al.  Analysis of the tertiary structure of protein β-sheet sandwiches , 1981 .

[22]  N. Xuong,et al.  Characteristics of a flat multiwire area detector for protein crystallography , 1981 .

[23]  J. Richardson,et al.  The anatomy and taxonomy of protein structure. , 1981, Advances in protein chemistry.

[24]  M. Belfort,et al.  A single functional arginyl residue involved in the catalysis promoted by Lactobacillus casei thymidylate synthetase. , 1980, Archives of biochemistry and biophysics.

[25]  H. G. Khorana,et al.  Amino acid sequence of bacteriorhodopsin. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[26]  K. Danenberg,et al.  Evidence for a sequential interaction of the subunits of thymidylate synthetase. , 1979, The Journal of biological chemistry.

[27]  A. V. Kiselev,et al.  The structural basis of the functioning of bacteriorhodopsin: An overview , 1979, FEBS letters.

[28]  G. Maley,et al.  The primary structure of Lactobacillus casei thymidylate synthetase. I. The isolation of cyanogen bromide peptides 1 through 5 and the complete amino acid sequence of CNBr 1, 2, 3, and 5. , 1979, The Journal of biological chemistry.

[29]  G. Maley,et al.  The primary structure of Lactobacillus casei thymidylate synthetase. II. The complete amino acid sequence of the active site peptide, CNBr 4. , 1979, The Journal of biological chemistry.

[30]  G. Maley,et al.  The primary structure of Lactobacillus casei thymidylate synthetase. III. The use of 2-(2-nitrophenylsulfenyl)-3-methyl-3-bromoindolenine and limited tryptic peptides to establish the complete amino acid sequence of the enzyme. , 1979, The Journal of biological chemistry.

[31]  H. G. Khorana.  Anderegg, R. J.,Nihei, K.,and Biemann Amino acid sequence of bacteriorhodopsin , 1979 .

[32]  W. A. Munroe,et al.  Effects of polyoxyanions on sulfhydryl group modification of thymidylate synthetase. , 1978, Biochemistry.

[33]  C. Blake,et al.  Do genes-in-pieces imply proteins-in-pieces? , 1978, Nature.

[34]  N. Xuong,et al.  The electron stationary picture method for high-speed measurement of reflection intensities from crystals with large unit cells , 1978 .

[35]  W. Gilbert.  Why genes in pieces? , 1978, Nature.

[36]  R. Stroud,et al.  Difference Fourier refinement of the structure of DIP‐trypsin at 1.5 Å with a minicomputer technique , 1977 .

[37]  Y. Wataya,et al.  Active-site labeling of thymidylate synthetase with 5-fluoro-2'-deoxyuridylate. , 1977, Methods in enzymology.

[38]  G. Bricogne,et al.  Methods and programs for direct‐space exploitation of geometric redundancies , 1976 .

[39]  J. Konnert,et al.  A restrained-parameter structure-factor least-squares refinement procedure for large asymmetric units , 1976 .

[40]  D. Santi,et al.  Thymidylate synthetase: studies on peptide containing covalently bound 5-fluoro-2'-deoxyuridylate and 5,10-methylenetetrahydrofolate. , 1976, Biochemical and biophysical research communications.

[41]  G. Maley,et al.  Amino acid sequence at the FdUMP binding site of thymidylate synthetase. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[42]  G. Maley,et al.  Factors affecting substrate binding in Lactobacillus casei thymidylate synthetase as studied by equilibrium dialysis. , 1976, Biochemistry.

[43]  R. Stroud,et al.  A pulsed diffusion technique for the growth of protein crystals for x-ray diffraction. , 1975, Journal of molecular biology.

[44]  C. Chothia,et al.  Principles of protein–protein recognition , 1975, Nature.

[45]  M. Caron,et al.  Identification of adenylate cyclase-coupled beta-adrenergic receptors in frog erythrocytes with (minus)-[3-H] alprenolol. , 1975, The Journal of biological chemistry.

[46]  J. Lyon,et al.  Gel electrophoresis as a means of detecting ternary complex formation of thymidylate synthetase , 1974 .

[47]  R. Dickerson,et al.  The structure of bovine trypsin : Electron density maps of the inhibited enzyme at 5 Å and at 2·7 Å resolution☆ , 1974 .

[48]  Cyrus Chothia,et al.  Conformation of twisted β-pleated sheets in proteins , 1973 .

[49]  B. Lee,et al.  The interpretation of protein structures: estimation of static accessibility. , 1971, Journal of molecular biology.

[50]  R. Leary,et al.  Properties of thymidylate synthetase from dichloromethotrexate-resistant Lactobacillus casei. , 1970, The Journal of biological chemistry.

[51]  E. Lattman,et al.  Representation of phase probability distributions for simplified combination of independent phase information , 1970 .

[52]  W. Fitch,et al.  EVOLUTIONARY VARIABILITY OF CYTOCHROME C PRIMARY STRUCTURES , 1968, Annals of the New York Academy of Sciences.

[53]  N. Allewell,et al.  Design of a diffractometer and flow cell system for X-ray analysis of crystalline proteins with applications to the crystal chemistry of ribonuclease-S. , 1967, Journal of molecular biology.

[54]  R. J. Swan,et al.  Optical rotatory dispersion of nucleic acid derivatives. 8. The conformation of pyrimidine nucleosides in solution. , 1967, Biochemistry.

[55]  J. C. Kendrew,et al.  The crystal structure of myoglobin: Phase determination to a resolution of 2 Å by the method of isomorphous replacement , 1961 .

[56]  G. A. Sim,et al.  The distribution of phase angles for structures containing heavy atoms. II. A modification of the normal heavy‐atom method for non‐centrosymmetrical structures , 1959 .

[57]  F. Richards,et al.  The Use of the Gradient Tube for the Determination of Crystal Densities1a , 1952 .