DOI: 10.1110/ps.17601. Understanding thermostability in cytochrome P450 by combinatorial mutagenesis

Understanding thermostability in cytochrome P450 by combinatorial mutagenesis

10.1110/ps.17601

Crossref journal-article

Wiley

Protein Science (311)

Abstract

AbstractThe 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.

Bibliography

Maves, S. A., & Sligar, S. G. (2001). Understanding thermostability in cytochrome P450 by combinatorial mutagenesis. Protein Science, 10(1), 161â168. Portico.

Authors 2

Shelley A. Maves (first)
Stephen G. Sligar (additional)

References 43 Referenced 36

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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 8 p.m.)
Deposited	1 year, 10 months ago (Oct. 8, 2023, 1:53 p.m.)
Indexed	1 year, 2 months ago (June 20, 2024, 5:13 p.m.)
Issued	24 years, 7 months ago (Jan. 1, 2001)
Published	24 years, 7 months ago (Jan. 1, 2001)
Published Online	16 years, 7 months ago (Dec. 31, 2008)
Published Print	24 years, 7 months ago (Jan. 1, 2001)

Funders 0

None

BibTeX

@article{Maves_2001, title={Understanding thermostability in cytochrome P450 by combinatorial mutagenesis}, volume={10}, ISSN={1469-896X}, url={http://dx.doi.org/10.1110/ps.17601}, DOI={10.1110/ps.17601}, number={1}, journal={Protein Science}, publisher={Wiley}, author={Maves, Shelley A. and Sligar, Stephen G.}, year={2001}, month=jan, pages={161–168} }

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