Q9XHE8 (C71DI_MENSP) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Cytochrome P450 71D18
EC=1.14.13.48

Alternative name(s):
(-)-(4S)-Limonene-6-hydroxylase

Gene names

Name:

CYP71D18

Organism

Mentha spicata (Spearmint)

Taxonomic identifier

29719 [NCBI]

Taxonomic lineage

Eukaryota › Viridiplantae › Streptophyta › Embryophyta › Tracheophyta › Spermatophyta › Magnoliophyta › eudicotyledons › core eudicotyledons › asterids › lamiids › Lamiales › Lamiaceae › Nepetoideae › Mentheae › Mentha

Protein attributes

Sequence length	496 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Hydroxylates (-)-(4S)-limonene to (-)-trans-carveol, a precursor of (-)-carvone, responsible for the typical spearmint flavor note. Fluorinated substrate analogs are hydroxylated with the same regio- and stereochemistry. Ref.1 Ref.3
Catalytic activity	--(S)-limonene + NADPH + O2 = (-)-trans-carveol + NADP+ + H2O. Ref.1 Ref.3
Cofactor	Heme group By similarity.
Subcellular location	Endoplasmic reticulum membrane; Single-pass type II membrane protein Potential.
Sequence similarities	Belongs to the cytochrome P450 family.

Ontologies

Keywords
Cellular component	Endoplasmic reticulum Membrane
Domain	Signal-anchor Transmembrane Transmembrane helix
Ligand	Heme Iron Metal-binding NADP
Molecular function	Monooxygenase Oxidoreductase
Technical term	Direct protein sequencing
Gene Ontology (GO)
Cellular component	endoplasmic reticulum membrane Inferred from electronic annotation. Source: UniProtKB-SubCell integral to membrane Inferred from electronic annotation. Source: UniProtKB-KW microsome Inferred from direct assay Ref.1. Source: UniProtKB
Molecular function	(S)-limonene 6-monooxygenase activity Inferred from direct assay Ref.1. Source: UniProtKB electron carrier activity Inferred from electronic annotation. Source: InterPro heme binding Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 496	496	Cytochrome P450 71D18		PRO_0000389497
Regions
	Transmembrane	2 – 22	21	Helical; Signal-anchor for type II membrane protein; Potential
Sites
	Metal binding	436	1	Iron (heme axial ligand) By similarity
Experimental info
	Mutagenesis	357	1	L → M: No effect on regiospecificity. Ref.2
	Mutagenesis	359	1	L → M: No effect on regiospecificity. Ref.2
	Mutagenesis	363	1	F → I: Conversion to the regiospecificity and catalytic efficiency of the peppermint limonene-3-hydroxylase. Ref.2
	Mutagenesis	369	1	Q → S: No effect on regiospecificity. Ref.2
	Mutagenesis	370	1	S → C: No effect on regiospecificity. Ref.2

Sequences

	Sequence		Length	Mass (Da)
	Q9XHE8 [UniParc]. Last modified November 1, 1999. Version 1. Checksum: 06E4370F95EB91C0	FASTA	496	56,149
	10 20 30 40 50 60 MELDLLSAII ILVATYIVSL LINQWRKSKS QQNLPPSPPK LPVIGHLHFL WGGLPQHVFR 70 80 90 100 110 120 SIAQKYGPVA HVQLGEVYSV VLSSAEAAKQ AMKVLDPNFA DRFDGIGSRT MWYDKDDIIF 130 140 150 160 170 180 SPYNDHWRQM RRICVTELLS PKNVRSFGYI RQEEIERLIR LLGSSGGAPV DVTEEVSKMS 190 200 210 220 230 240 CVVVCRAAFG SVLKDQGSLA ELVKESLALA SGFELADLYP SSWLLNLLSL NKYRLQRMRR 250 260 270 280 290 300 RLDHILDGFL EEHREKKSGE FGGEDIVDVL FRMQKGSDIK IPITSNCIKG FIFDTFSAGA 310 320 330 340 350 360 ETSSTTISWA LSELMRNPAK MAKVQAEVRE ALKGKTVVDL SEVQELKYLR SVLKETLRLH 370 380 390 400 410 420 PPFPLIPRQS REECEVNGYT IPAKTRIFIN VWAIGRDPQY WEDPDTFRPE RFDEVSRDFM 430 440 450 460 470 480 GNDFEFIPFG AGRRICPGLH FGLANVEIPL AQLLYHFDWK LPQGMTDADL DMTETPGLSG 490 PKKKNVCLVP TLYKSP

References

[1]	"Regiospecific cytochrome P450 limonene hydroxylases from mint (Mentha) species: cDNA isolation, characterization, and functional expression of (-)-4S-limonene-3-hydroxylase and (-)-4S-limonene-6-hydroxylase." Lupien S., Karp F., Wildung M., Croteau R. Arch. Biochem. Biophys. 368:181-192(1999) [PubMed: 10415126] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 1-21; 176-200 AND 375-398, CATALYTIC ACTIVITY, FUNCTION.
[2]	"A single amino acid substitution (F363I) converts the regiochemistry of the spearmint (-)-limonene hydroxylase from a C6- to a C3-hydroxylase." Schalk M., Croteau R. Proc. Natl. Acad. Sci. U.S.A. 97:11948-11953(2000) [PubMed: 11050228] [Abstract] Cited for: MUTAGENESIS OF LEU-357; LEU-359; PHE-363; GLN-369 AND SER-370.
[3]	"Hydroxylation of limonene enantiomers and analogs by recombinant (-)-limonene 3- and 6-hydroxylases from mint (Mentha) species: evidence for catalysis within sterically constrained active sites." Wuest M., Little D.B., Schalk M., Croteau R. Arch. Biochem. Biophys. 387:125-136(2001) [PubMed: 11368174] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY.

Web resources

Protein Spotlight

Mint condition - Issue 113 of January 2010

Cross-references

Sequence databases
EMBL GenBank DDBJ	AF124815 mRNA. Translation: AAD44150.1.
3D structure databases
HSSP	HSSP built from PDB template 1YQP based on UniProtKB P14779.
ProteinModelPortal	Q9XHE8.
ModBase	Search...
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Enzyme and pathway databases
BioCyc	MetaCyc:MONOMER-15424.
Family and domain databases
InterPro	IPR001128. Cyt_P450. IPR017972. Cyt_P450_CS. IPR002401. Cyt_P450_E_grp-I. [Graphical view]
Gene3D	G3DSA:1.10.630.10. Cyt_P450. 1 hit.
Pfam	PF00067. p450. 1 hit. [Graphical view]
PRINTS	PR00463. EP450I. PR00385. P450.
SUPFAM	SSF48264. Cytochrome_P450. 1 hit.
PROSITE	PS00086. CYTOCHROME_P450. 1 hit. [Graphical view]
ProtoNet	Search...

Entry information

Entry name

C71DI_MENSP

Accession

Primary (citable) accession number: Q9XHE8

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 24, 2009
Last sequence update:	November 1, 1999
Last modified:	September 21, 2011
This is version 56 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Plant Protein Annotation Program