Q03471 (ARIS_PENRO) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Aristolochene synthase
Short name=AS
EC=4.2.3.9

Alternative name(s):
Sesquiterpene cyclase

Gene names

Name:

ARI1

Organism

Penicillium roqueforti

Taxonomic identifier

5082 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Fungi › Dikarya › Ascomycota › saccharomyceta › Pezizomycotina › leotiomyceta › Eurotiomycetes › Eurotiomycetidae › Eurotiales › Trichocomaceae › mitosporic Trichocomaceae › Penicillium

Protein attributes

Sequence length	342 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the cyclization of trans,trans-farnesyl diphosphate (FPP) to the bicyclic sesquiterpene aristolochene. Produces germacrene A as an enzyme-bound intermediate that is not released by the enzyme, but is further cyclized to produce aristolochene. Aristolochene is the likely parent compound for a number of sesquiterpenoid toxins produced by filamentous fungi. Ref.1 Ref.2
Catalytic activity	(2E,6E)-farnesyl diphosphate = aristolochene + diphosphate. Ref.1
Cofactor	Binds 3 magnesium ions per subunit By similarity.
Pathway	Sesquiterpene biosynthesis; aristolochene biosynthesis; aristolochene from farnesyl diphosphate: step 1/1.
Subunit structure	Homodimer By similarity.
Subcellular location	Cytoplasm.
Sequence similarities	Belongs to the terpene synthase family.
Biophysicochemical properties	Kinetic parameters: KM=0.6 µM for (2E,6E)-farnesyl diphosphate Ref.2

Ontologies

Keywords
Cellular component	Cytoplasm
Ligand	Magnesium Metal-binding
Molecular function	Lyase
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	aristolochene synthase activity Inferred from electronic annotation. Source: EC magnesium ion 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 – 342

342

Aristolochene synthase

PRO_0000064677

Regions

Region

340 – 341

2

Substrate-binding By similarity

Sites

Metal binding

115

1

Magnesium 1 By similarity

Metal binding

115

1

Magnesium 2 By similarity

Metal binding

244

1

Magnesium 3 By similarity

Metal binding

248

1

Magnesium 3 By similarity

Metal binding

252

1

Magnesium 3 By similarity

Binding site

200

1

Substrate By similarity

Binding site

251

1

Substrate By similarity

Experimental info

Mutagenesis

92

1

Y → F: Causes 100-fold reduction in kcat but a 50-fold decrease in KM, resulting in a 2-fold decrease in catalytic efficiency. Ref.2

Mutagenesis

115

1

D → N: Abolishes catalytic activity. Ref.2

Mutagenesis

244

1

N → L: Abolishes catalytic activity. Ref.2

Mutagenesis

248

1

S → A: Abolishes catalytic activity; when associated with D-252. Ref.2

Mutagenesis

252

1

E → D: Abolishes catalytic activity; when associated with A-248. Ref.2

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

342

Helix Strand Turn

Details...

Helix

54 – 68

15

Helix

74 – 83

10

Helix

85 – 92

8

Turn

98 – 100

3

Helix

101 – 120

20

Helix

123 – 137

15

Helix

148 – 180

33

Turn

181 – 183

3

Helix

193 – 201

9

Helix

204 – 217

14

Helix

223 – 227

5

Helix

230 – 248

19

Turn

249 – 252

4

Helix

269 – 277

9

Helix

281 – 307

27

Helix

315 – 336

22

Sequences

	Sequence		Length	Mass (Da)
	Q03471 [UniParc]. Last modified October 1, 1993. Version 1. Checksum: ACFBD63F4FF9BB03	FASTA	342	39,192
	10 20 30 40 50 60 MATSTETISS LAQPFVHLEN PINSPLVKET IRPRNDTTIT PPPTQWSYLC HPRVKEVQDE 70 80 90 100 110 120 VDGYFLENWK FPSFKAVRTF LDAKFSEVTC LYFPLALDDR IHFACRLLTV LFLIDDVLEH 130 140 150 160 170 180 MSFADGEAYN NRLIPISRGD VLPDRTKPEE FILYDLWESM RAHDAELANE VLEPTFVFMR 190 200 210 220 230 240 AQTDRARLSI HELGHYLEYR EKDVGKALLS ALMRFSMGLR LSADELQDMK ALEANCAKQL 250 260 270 280 290 300 SVVNDIYSYD KEEEASRTGH KEGAFLCSAV KVLAEESKLG IPATKRVLWS MTREWETVHD 310 320 330 340 EIVAEKIASP DGCSEAAKAY MKGLEYQMSG NEQWSKTTRR YN

References

[1]	"Aristolochene synthase. Isolation, characterization, and bacterial expression of a sesquiterpenoid biosynthetic gene (Ari1) from Penicillium roqueforti." Proctor R.H., Hohn T.M. J. Biol. Chem. 268:4543-4548(1993) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 56-70; 76-84; 245-251 AND 323-336, FUNCTION, CATALYTIC ACTIVITY. Strain: ATCC 10110 / CBS 221.30 / NBRC 5459 / NRRL 849.
[2]	"Aristolochene synthase: mechanistic analysis of active site residues by site-directed mutagenesis." Felicetti B., Cane D.E. J. Am. Chem. Soc. 126:7212-7221(2004) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF TYR-92; ASP-115; ASN-244; SER-248 AND GLU-252.
[3]	"Crystal structure determination of aristolochene synthase from the blue cheese mold, Penicillium roqueforti." Caruthers J.M., Kang I., Rynkiewicz M.J., Cane D.E., Christianson D.W. J. Biol. Chem. 275:25533-25539(2000) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS).

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

L05193 Genomic DNA. Translation: AAA33694.1.

PIR

A45462.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1DGP	X-ray	2.80	A/B	40-339	[»]
1DI1	X-ray	2.50	A/B	40-339	[»]
1F1K	model	-	A	1-342	[»]
1F1L	model	-	A	1-342	[»]
1F1N	model	-	A	1-342	[»]
1F1P	model	-	A	1-342	[»]

ProteinModelPortal

Q03471.

SMR

Q03471. Positions 40-339.

ModBase

Search...

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Enzyme and pathway databases

BioCyc

MetaCyc:MONOMER-16547.

BRENDA

4.2.3.9. 8737.

UniPathway

UPA00177; UER00582.

Family and domain databases

Gene3D

1.10.600.10. 1 hit.

InterPro

IPR005630. Terpene_synthase_metal-bd.
IPR008949. Terpenoid_synth.
[Graphical view]

Pfam

PF03936. Terpene_synth_C. 1 hit.
[Graphical view]

SUPFAM

SSF48576. Terpenoid_synth. 1 hit.

ProtoNet

Search...

Other

EvolutionaryTrace

Q03471.

Entry information

Entry name

ARIS_PENRO

Accession

Primary (citable) accession number: Q03471

Entry history

Integrated into UniProtKB/Swiss-Prot:	October 1, 1993
Last sequence update:	October 1, 1993
Last modified:	April 3, 2013
This is version 82 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Fungal Protein Annotation Program