The cytochrome P450 gene family CYP157 does not contain EXXR in the K-helix reducing the absolute conserved P450 residues to a single cysteine

Sanjeewa Rupasinghe; Mary A. Schuler; Norio Kagawa; Hang Yuan; Li Lei; Bin Zhao; Steven L. Kelly; Michael R. Waterman; David C. Lamb

doi:10.1016/j.febslet.2006.10.043

The cytochrome P450 gene family CYP157 does not contain EXXR in the K-helix reducing the absolute conserved P450 residues to a single cysteine

Sanjeewa Rupasinghe, Mary A. Schuler, Norio Kagawa, Hang Yuan, Li Lei, Bin Zhao, Steven L. Kelly, Michael R. Waterman, David C. Lamb

Research output: Contribution to journal › Article

Abstract

In this work, we have spectroscopically characterised CYP157C1 from Streptomyces coelicolor A3(2) which has the motif E²⁹⁷QSLW³⁰¹ rather than the invariant EXXR motif in the P450 K-helix. Site-directed mutagenesis of native E²⁹⁷QSLW³⁰¹ in CYP157C1 to E²⁹⁷ESLR³⁰¹ or E²⁹⁷QSRW³⁰¹ both containing standard EXXR motifs produced cytochrome P420 proteins thought to be inactive forms of P450 even though wild type CYP157C1 has the spectral properties of a normal P450. These results indicate that the EXXR motif is not required in all CYP tertiary architectures and only a single cysteine residue, which coordinates as the fifth thiolate ligand to the P450 haem iron, is invariant in all CYPs structures.

Original language	English (US)
Pages (from-to)	6338-6342
Number of pages	5
Journal	FEBS Letters
Volume	580
Issue number	27
DOIs	https://doi.org/10.1016/j.febslet.2006.10.043
State	Published - Nov 27 2006

Keywords

Cytochrome P450
Protein folding
Site-directed mutagenesis
Streptomyces

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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Rupasinghe, S., Schuler, M. A., Kagawa, N., Yuan, H., Lei, L., Zhao, B., Kelly, S. L., Waterman, M. R., & Lamb, D. C. (2006). The cytochrome P450 gene family CYP157 does not contain EXXR in the K-helix reducing the absolute conserved P450 residues to a single cysteine. FEBS Letters, 580(27), 6338-6342. https://doi.org/10.1016/j.febslet.2006.10.043

The cytochrome P450 gene family CYP157 does not contain EXXR in the K-helix reducing the absolute conserved P450 residues to a single cysteine. / Rupasinghe, Sanjeewa; Schuler, Mary A.; Kagawa, Norio; Yuan, Hang; Lei, Li; Zhao, Bin; Kelly, Steven L.; Waterman, Michael R.; Lamb, David C.

In: FEBS Letters, Vol. 580, No. 27, 27.11.2006, p. 6338-6342.

Research output: Contribution to journal › Article

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abstract = "In this work, we have spectroscopically characterised CYP157C1 from Streptomyces coelicolor A3(2) which has the motif E297QSLW301 rather than the invariant EXXR motif in the P450 K-helix. Site-directed mutagenesis of native E297QSLW301 in CYP157C1 to E297ESLR301 or E297QSRW301 both containing standard EXXR motifs produced cytochrome P420 proteins thought to be inactive forms of P450 even though wild type CYP157C1 has the spectral properties of a normal P450. These results indicate that the EXXR motif is not required in all CYP tertiary architectures and only a single cysteine residue, which coordinates as the fifth thiolate ligand to the P450 haem iron, is invariant in all CYPs structures.",

keywords = "Cytochrome P450, Protein folding, Site-directed mutagenesis, Streptomyces",

author = "Sanjeewa Rupasinghe and Schuler, {Mary A.} and Norio Kagawa and Hang Yuan and Li Lei and Bin Zhao and Kelly, {Steven L.} and Waterman, {Michael R.} and Lamb, {David C.}",

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AU - Rupasinghe, Sanjeewa

AU - Schuler, Mary A.

AU - Kagawa, Norio

AU - Yuan, Hang

AU - Lei, Li

AU - Zhao, Bin

AU - Kelly, Steven L.

AU - Waterman, Michael R.

AU - Lamb, David C.

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AB - In this work, we have spectroscopically characterised CYP157C1 from Streptomyces coelicolor A3(2) which has the motif E297QSLW301 rather than the invariant EXXR motif in the P450 K-helix. Site-directed mutagenesis of native E297QSLW301 in CYP157C1 to E297ESLR301 or E297QSRW301 both containing standard EXXR motifs produced cytochrome P420 proteins thought to be inactive forms of P450 even though wild type CYP157C1 has the spectral properties of a normal P450. These results indicate that the EXXR motif is not required in all CYP tertiary architectures and only a single cysteine residue, which coordinates as the fifth thiolate ligand to the P450 haem iron, is invariant in all CYPs structures.

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