Understanding thermostability in cytochrome P450 by combinatorial mutagenesis

Shelley A. Maves, Stephen G. Sligar

Research output: Contribution to journalArticlepeer-review


The cytochromes P450 are an important class of mono-oxygenases involved in xenobiotic metabolism and steroid biosynthesis in a diverse set of life forms. Discovery of CYP-119, a P450 from the archea Sulfolobus solfataricus has provided a means for understanding nature's method of stabilizing this important protein superfamily. To identify classes of stabilizing interactions used by CYP-119, we have generated a randomized library of point mutants and screened for mutants that are less thermostable than the wild type by monitoring the characteristic Soret band in the visible region of the cell lysis. The selected mutants were characterized by differential scanning calorimetry to compare the temperatures of the melting transitions of the various mutants. The identified mutations suggested that electrostatic interactions involving salt links and charge-charge interactions, as well as contributions from other interactions such as aromatic stacking, and side chain volume of hydrophobic residues contribute to enhanced thermostability in this cytochrome P450.

Original languageEnglish (US)
Pages (from-to)161-168
Number of pages8
JournalProtein Science
Issue number1
StatePublished - 2001


  • Cytochrome P450
  • Electrostatic interactions
  • Random mutagenesis
  • Salt links
  • Thermostability

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


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