Processo Para Determinação Pós-genèmica De Estruturas Atèmicas Macromoleculares

  • Published: Apr 9, 2002
  • Earliest Priority: Jan 22 1999
  • Family: 11
  • Cited Works: 0
  • Cited by: 0
  • Cites: 0
  • Additional Info: Published
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Abstract

A process for pan-genomic determination of three-dimensional macromolecular atomic structures uses a unique combination of components. All known structural information, sequence information and functional information are systematically organized into a genomics database. Advanced tools of bioinformatics are used to cluster all known gene products into families of homologous sequences. Simultaneously, in parallel for each such family, a few cDNAs from appropriately representatives species are cloned into expression vectors for a few expressions systems. Constructs are then screened for expression, and those that are effective advance to the preparative step. Expressed proteins are prepared, purified and characterized. Purified proteins are set to crystallize in parallel against crystallization screens. Crystals that grow are tested for suitable diffraction characteristics. A suitable crystal is frozen, and diffraction data are measured using the multiwavelength anomalous diffraction (MAD) method at a synchrotron which uses undulator beamlines for high-throughput crystallography. Diffraction data are analyzed by the MAD phasing method, an atomic model is built, and the model is refined against the diffraction data. The refined model is analyzed in the context of (1) sequence information from other family members, (2) all other known 3D structures, and (3) functional motifs. It is also analyzed for surface characteristics with the aim to define active sites and macromolecular contact sites. For relevant structures, the active site properties are used to define classes of compounds predicted to have binding potency. Computational tools for homology model building are used to develop models for homologs. The homology models may be used in target selection, drug design, or design of more appropriate constructs for experimental analysis. The ensemble of all known structures is used to further advance the effectiveness of the bioinformatics tools.


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Document History
  • Publication: Apr 9, 2002
  • Application: Jan 21, 2000
    BR BR 0007638 A
  • Priority: Jan 21, 2000
    US US 0001600 W
  • Priority: Jan 22, 1999
    US US 23598699 A

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