Characterization of four orthologs of stringent starvation protein A

Acta Crystallogr D Biol Crystallogr. 2003 May;59(Pt 5):881-6. doi: 10.1107/s0907444903005699. Epub 2003 Apr 25.

Abstract

Orthologous proteins can be beneficial for X-ray crystallographic studies when a protein from an organism of choice fails to crystallize or the crystals are not suitable for structure determination. Their amino-acid sequences should be similar enough that they will share the same fold, but different enough so that they may crystallize under alternative conditions and diffract to higher resolution. This multi-species approach was employed to obtain diffraction-quality crystals of the RNA polymerase (RNAP) associated stringent starvation protein A (SspA). Although Escherichia coli SspA could be crystallized, the crystals failed to diffract well enough for structure determination. Therefore, SspA proteins from Yersinia pestis, Vibrio cholerae and Pseudomonas aeruginosa were cloned, expressed, purified and subjected to crystallization trials. The V. cholerae SspA protein failed to crystallize under any conditions tested and the P. aeruginosa SspA protein did not form crystals suitable for data collection. On the other hand, Y. pestis SspA crystallized readily and the crystals diffracted to 2.0 A.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Cloning, Molecular
  • Crystallization
  • Escherichia coli / chemistry
  • Escherichia coli / genetics
  • Escherichia coli Proteins / biosynthesis
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / genetics
  • Molecular Sequence Data
  • Pseudomonas aeruginosa / chemistry
  • Pseudomonas aeruginosa / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Vibrio cholerae / chemistry
  • Vibrio cholerae / genetics
  • X-Ray Diffraction
  • Yersinia pestis / chemistry
  • Yersinia pestis / genetics

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • Recombinant Proteins
  • SspA protein, E coli