Abstract
In this work, we have spectroscopically characterised CYP157C1 from Streptomyces coelicolor A3(2) which has the motif E 297 QSLW 301 rather than the invariant EXXR motif in the P450 K-helix. Site-directed mutagenesis of native E 297 QSLW 301 in CYP157C1 to E 297 ESLR 301 or E 297 QSRW 301 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 | |
| State | Published - Nov 27 2006 |
Fingerprint
Keywords
- Cytochrome P450
- Protein folding
- Site-directed mutagenesis
- Streptomyces
ASJC Scopus subject areas
- Biophysics
- Structural Biology
- Biochemistry
- Molecular Biology
- Genetics
- Cell Biology
Cite this
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
}
TY - JOUR
T1 - The cytochrome P450 gene family CYP157 does not contain EXXR in the K-helix reducing the absolute conserved P450 residues to a single cysteine
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.
PY - 2006/11/27
Y1 - 2006/11/27
N2 - In this work, we have spectroscopically characterised CYP157C1 from Streptomyces coelicolor A3(2) which has the motif E 297 QSLW 301 rather than the invariant EXXR motif in the P450 K-helix. Site-directed mutagenesis of native E 297 QSLW 301 in CYP157C1 to E 297 ESLR 301 or E 297 QSRW 301 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.
AB - In this work, we have spectroscopically characterised CYP157C1 from Streptomyces coelicolor A3(2) which has the motif E 297 QSLW 301 rather than the invariant EXXR motif in the P450 K-helix. Site-directed mutagenesis of native E 297 QSLW 301 in CYP157C1 to E 297 ESLR 301 or E 297 QSRW 301 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.
KW - Cytochrome P450
KW - Protein folding
KW - Site-directed mutagenesis
KW - Streptomyces
UR - http://www.scopus.com/inward/record.url?scp=33750992460&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33750992460&partnerID=8YFLogxK
U2 - 10.1016/j.febslet.2006.10.043
DO - 10.1016/j.febslet.2006.10.043
M3 - Article
C2 - 17092500
AN - SCOPUS:33750992460
VL - 580
SP - 6338
EP - 6342
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 27
ER -