BIOMAT Database Logo BIOMAT Database Logo

Crystallization and preliminary X-ray analysis of a DNA primase from hyperthermophilic archaeon Pyrococcus horikoshii. 2001

N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Cellular Physiology Laboratory, RIKEN (The Institute of Physical and Chemical Research), 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

At the initiation of chromosomal DNA replication, DNA primases synthesize short RNA primers, which are subsequently elongated by DNA polymerases. To understand the structural basis for the primer synthesis by archaeal/eukaryotic-type primases, the gene of the DNA primase from hyperthermophilic archaeon Pyrococcus horikoshii was cloned and overexpressed in Escherichia coli as a fusion protein with a hexa-histidine tag at its amino terminus. The recombinant DNA primase was purified and crystallized by the hanging-drop vapor diffusion method at 293 K, with polyethylene glycol 8000 as the precipitant. The crystals belong to the P3(2)21 space group with unit-cell parameters a = b = 77.8, c = 129.6 A, and alpha = beta = 90 degrees, gamma = 120 degrees. Crystals of the selenomethionine derivative were obtained by means of a cross-seeding method using native crystals. The data for the native and selenomethionine-substituted crystals were collected to 1.8 and 2.2 A resolution, respectively, with synchrotron radiation at SPring-8 under flash-frozen conditions at 100 K. The four wavelength MAD data provided a phase to determine the structure of the primase at 2.2 A resolution.

UI MeSH Term Description Entries
D011487 Protein Conformation The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain). Conformation, Protein,Conformations, Protein,Protein Conformations
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D003460 Crystallization The formation of crystalline substances from solutions or melts. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Crystalline Polymorphs,Polymorphism, Crystallization,Crystal Growth,Polymorphic Crystals,Crystal, Polymorphic,Crystalline Polymorph,Crystallization Polymorphism,Crystallization Polymorphisms,Crystals, Polymorphic,Growth, Crystal,Polymorph, Crystalline,Polymorphic Crystal,Polymorphisms, Crystallization,Polymorphs, Crystalline
D012313 RNA A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed) RNA, Non-Polyadenylated,Ribonucleic Acid,Gene Products, RNA,Non-Polyadenylated RNA,Acid, Ribonucleic,Non Polyadenylated RNA,RNA Gene Products,RNA, Non Polyadenylated
D012645 Selenomethionine Diagnostic aid in pancreas function determination. Butanoic acid, 2-amino-4-(methylseleno)-,Radioselenomethionine,Selenomethionine Se 75,Selenomethionine Hydrochloride, (S)-Isomer,Selenomethionine, (+,-)-Isomer,Selenomethionine, (R)-Isomer,Selenomethionine, (S)-Isomer,Sethotope,Se 75, Selenomethionine
D018360 Crystallography, X-Ray The study of crystal structure using X-RAY DIFFRACTION techniques. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) X-Ray Crystallography,Crystallography, X Ray,Crystallography, Xray,X Ray Crystallography,Xray Crystallography,Crystallographies, X Ray,X Ray Crystallographies
D019711 Pyrococcus A genus of strictly anaerobic ultrathermophilic archaea, in the family THERMOCOCCACEAE, occurring in heated seawaters. They exhibit heterotrophic growth at an optimum temperature of 100 degrees C.
D019915 DNA Primase A single-stranded DNA-dependent RNA polymerase that functions to initiate, or prime, DNA synthesis by synthesizing oligoribonucleotide primers. EC 2.7.7.-. Primase,Bacteriophage T7 Gene 4 Protein,DnaG (Primase),DnaG Gene Product,DnaG Protein,T7 DNA Primase-Helicase Protein,T7 DNA-Priming Protein,T7 gene-4 protein,DNA-Priming Protein, T7,Primase, DNA,Protein, T7 DNA-Priming,T7 DNA Primase Helicase Protein,T7 DNA Priming Protein,T7 gene 4 protein
D019943 Amino Acid Substitution The naturally occurring or experimentally induced replacement of one or more AMINO ACIDS in a protein with another. If a functionally equivalent amino acid is substituted, the protein may retain wild-type activity. Substitution may also diminish, enhance, or eliminate protein function. Experimentally induced substitution is often used to study enzyme activities and binding site properties. Amino Acid Substitutions,Substitution, Amino Acid,Substitutions, Amino Acid

Related Publications

N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
The MinD protein from the hyperthermophilic archaeon Pyrococcus horikoshii: crystallization and preliminary X-ray analysis.
June 2001, Acta crystallographica. Section D, Biological crystallography,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Crystallization and preliminary X-ray analysis of inorganic pyrophosphatase from the hyperthermophilic archaeon Pyrococcus horikoshii OT3.
March 2004, Acta crystallographica. Section D, Biological crystallography,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Crystallization and preliminary X-ray analysis of hyperthermophilic L-threonine dehydrogenase from the archaeon Pyrococcus horikoshii.
April 2005, Acta crystallographica. Section F, Structural biology and crystallization communications,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Crystallization and preliminary X-ray analysis of recombinant histone HPhA from the hyperthermophilic archaeon Pyrococcus horikoshii OT3.
May 2002, Acta crystallographica. Section D, Biological crystallography,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Crystallization and preliminary crystallographic analysis of an acylphosphatase from the hyperthermophilic archaeon Pyrococcus horikoshii.
July 2004, Acta crystallographica. Section D, Biological crystallography,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Crystallization and preliminary X-ray analysis of PH1566, a putative ribosomal RNA-processing factor from the hyperthermophilic archaeon Pyrococcus horikoshii OT3.
January 2006, Acta crystallographica. Section F, Structural biology and crystallization communications,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Expression, purification, crystallization and preliminary X-ray analysis of the KaiC-like protein PH0187 from the hyperthermophilic archaeon Pyrococcus horikoshii OT3.
January 2011, Acta crystallographica. Section F, Structural biology and crystallization communications,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Crystallization and preliminary X-ray analysis of endoglucanase from Pyrococcus horikoshii.
December 2008, Acta crystallographica. Section F, Structural biology and crystallization communications,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Crystallization and preliminary X-ray crystallographic study of leucyl-tRNA synthetase from the archaeon Pyrococcus horikoshii.
October 2004, Acta crystallographica. Section D, Biological crystallography,
N Ito, and O Nureki, and M Shirouzu, and S Yokoyama, and F Hanaoka
Crystallization and preliminary X-ray study of Pk--REC from a hyperthermophilic archaeon, Pyrococcus kodakaraensis KOD1.
May 2000, Acta crystallographica. Section D, Biological crystallography,
Copied contents to your clipboard!