Q40577 (5EAS_TOBAC) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
5-epi-aristolochene synthase
Short name=EAS
EC=4.2.3.61

Gene names

Name:	EAS3
AND
Name:	EAS4

Organism

Nicotiana tabacum (Common tobacco)

Taxonomic identifier

4097 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Viridiplantae › Streptophyta › Streptophytina › Embryophyta › Tracheophyta › Euphyllophyta › Spermatophyta › Magnoliophyta › eudicotyledons › core eudicotyledons › asterids › lamiids › Solanales › Solanaceae › Nicotianoideae › Nicotianeae › Nicotiana

Protein attributes

Sequence length	548 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 intermediate 5-epi-aristolochene, initial step in the conversion of FPP to the sesquiterpenoid antifungal phytoalexin capsidiol. Produces germacrene A as an enzyme-bound intermediate that is not released by the enzyme, but is further cyclized to produce the bicyclic 5-epi-aristolochene.
Catalytic activity	(2E,6E)-farnesyl diphosphate = (+)-5-epiaristolochene + diphosphate. Ref.3
Cofactor	Binds 3 magnesium ions per subunit.
Pathway	Secondary metabolite biosynthesis; terpenoid biosynthesis.
Subunit structure	Monomer.
Subcellular location	Cytoplasm.
Induction	By fungal elicitor. Ref.1
Domain	The Asp-Asp-Xaa-Xaa-Asp/Glu (DDXXD/E) motif is important for the catalytic activity, presumably through binding to Mg2+.
Miscellaneous	Ref.1 indicates the presence of at least two genes encoding for the same protein.
Sequence similarities	Belongs to the terpene synthase family.
Biophysicochemical properties	Kinetic parameters: KM=2.3 µM for 2-trans,6-trans-farnesyl diphosphate Ref.3

Ontologies

Keywords
Cellular component	Cytoplasm
Ligand	Magnesium Metal-binding
Molecular function	Lyase
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological_process	terpenoid biosynthetic process Inferred from electronic annotation. Source: UniProtKB-UniPathway
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	magnesium ion binding Inferred from electronic annotation. Source: InterPro terpene synthase activity 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 – 548

548

5-epi-aristolochene synthase

PRO_0000186438

Regions

Motif

301 – 305

5

DDXXD motif

Sites

Metal binding

301

1

Magnesium 1

Metal binding

301

1

Magnesium 2

Metal binding

305

1

Magnesium 1

Metal binding

305

1

Magnesium 2

Metal binding

444

1

Magnesium 3

Metal binding

448

1

Magnesium 3

Metal binding

452

1

Magnesium 3

Experimental info

Mutagenesis

520

1

Y → F: Loss of production of aristolochene, and accumulation of the intermediate germacrene A. Ref.3

Sequence conflict

35

1

K → D in AAA19216. Ref.1

Sequence conflict

42

1

Y → YIY AA sequence Ref.1

Sequence conflict

44

1

K → Q in AAA19216. Ref.1

Sequence conflict

55

1

N → S in AAA19216. Ref.1

Sequence conflict

62

1

M → R in AAA19216. Ref.1

Sequence conflict

73

1

T → I in AAA19216. Ref.1

Sequence conflict

89

1

D → E in AAA19216. Ref.1

Sequence conflict

388

1

T → M in AAA19216. Ref.1

Secondary structure

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548

Helix Strand Turn

Details...

Turn

24 – 29

6

Helix

36 – 57

22

Helix

64 – 76

13

Helix

80 – 82

3

Helix

84 – 97

14

Helix

104 – 116

13

Helix

123 – 129

7

Beta strand

134 – 136

3

Helix

138 – 142

5

Helix

144 – 154

11

Helix

162 – 164

3

Helix

167 – 178

12

Helix

179 – 181

3

Helix

186 – 195

10

Helix

203 – 213

11

Helix

215 – 217

3

Helix

223 – 252

30

Turn

253 – 255

3

Helix

256 – 259

4

Helix

267 – 277

11

Helix

281 – 283

3

Helix

284 – 305

22

Helix

310 – 322

13

Helix

325 – 330

6

Helix

333 – 354

22

Turn

355 – 357

3

Helix

359 – 361

3

Helix

362 – 385

24

Helix

391 – 398

8

Helix

400 – 402

3

Helix

404 – 413

10

Helix

420 – 427

8

Helix

431 – 447

17

Helix

449 – 454

6

Helix

461 – 469

9

Helix

473 – 494

22

Beta strand

496 – 498

3

Helix

503 – 505

3

Helix

507 – 519

13

Turn

520 – 522

3

Turn

523 – 525

3

Turn

526 – 528

3

Helix

531 – 542

12

Sequences

	Sequence		Length	Mass (Da)
	Q40577 [UniParc]. Last modified April 29, 2008. Version 3. Checksum: 9FE25FF361A68BF1	FASTA	548	62,987
	10 20 30 40 50 60 MASAAVANYE EEIVRPVADF SPSLWGDQFL SFSIKNQVAE KYAKEIEALK EQTRNMLLAT 70 80 90 100 110 120 GMKLADTLNL IDTIERLGIS YHFEKEIDDI LDQIYNQNSN CNDLCTSALQ FRLLRQHGFN 130 140 150 160 170 180 ISPEIFSKFQ DENGKFKESL ASDVLGLLNL YEASHVRTHA DDILEDALAF STIHLESAAP 190 200 210 220 230 240 HLKSPLREQV THALEQCLHK GVPRVETRFF ISSIYDKEQS KNNVLLRFAK LDFNLLQMLH 250 260 270 280 290 300 KQELAQVSRW WKDLDFVTTL PYARDRVVEC YFWALGVYFE PQYSQARVML VKTISMISIV 310 320 330 340 350 360 DDTFDAYGTV KELEAYTDAI QRWDINEIDR LPDYMKISYK AILDLYKDYE KELSSAGRSH 370 380 390 400 410 420 IVCHAIERMK EVVRNYNVES TWFIEGYTPP VSEYLSNALA TTTYYYLATT SYLGMKSATE 430 440 450 460 470 480 QDFEWLSKNP KILEASVIIC RVIDDTATYE VEKSRGQIAT GIECCMRDYG ISTKEAMAKF 490 500 510 520 530 540 QNMAETAWKD INEGLLRPTP VSTEFLTPIL NLARIVEVTY IHNLDGYTHP EKVLKPHIIN LLVDSIKI

References

[1]	"Gene family for an elicitor-induced sesquiterpene cyclase in tobacco." Facchini P.J., Chappell J. Proc. Natl. Acad. Sci. U.S.A. 89:11088-11092(1992) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA / MRNA], PROTEIN SEQUENCE OF 56-73, INDUCTION. Strain: cv. NK 326.
[2]	O'Maille P.E. Unpublished observations (APR-2008) Cited for: SEQUENCE REVISION TO 35.
[3]	"Demonstration of germacrene A as an intermediate in 5-epi-aristolochene synthase catalysis." Rising K.A., Starks C.M., Noel J.P., Chappell J. J. Am. Chem. Soc. 122:1861-1866(2000) Cited for: CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF TYR-520.
[4]	"Structural basis for cyclic terpene biosynthesis by tobacco 5-epi-aristolochene synthase." Starks C.M., Back K., Chappell J., Noel J.P. Science 277:1815-1820(1997) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS) IN COMPLEX WITH MAGNESIUM IONS AND SUBSTRATE, SEQUENCE REVISION.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

L04680 Unassigned RNA. Translation: AAA19216.1.

PIR

T03714.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1HX9	X-ray	3.50	A	1-548	[»]
1HXA	X-ray	2.32	A	1-548	[»]
1HXC	X-ray	2.25	A	1-548	[»]
1HXG	X-ray	2.90	A	1-548	[»]
3LZ9	X-ray	2.28	A	1-548	[»]
3M00	X-ray	2.10	A	1-548	[»]
3M01	X-ray	2.60	A	1-548	[»]
3M02	X-ray	2.50	A	1-548	[»]
4DI5	X-ray	2.30	A	14-548	[»]
5EAS	X-ray	2.25	A	1-548	[»]
5EAT	X-ray	2.80	A	1-548	[»]
5EAU	X-ray	2.15	A	1-548	[»]

ProteinModelPortal

Q40577.

SMR

Q40577. Positions 21-548.

ModBase

Search...

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Enzyme and pathway databases

BioCyc

MetaCyc:EAS-MONOMER.

BRENDA

4.2.3.9. 3645.

UniPathway

UPA00213.

Family and domain databases

Gene3D

1.10.600.10. 1 hit.
1.50.30.10. 1 hit.

InterPro

IPR001906. Terpene_synth_N.
IPR005630. Terpene_synthase_metal-bd.
IPR008930. Terpenoid_cyclase/PrenylTrfase.
IPR008949. Terpenoid_synth.
[Graphical view]

Pfam

PF01397. Terpene_synth. 1 hit.
PF03936. Terpene_synth_C. 1 hit.
[Graphical view]

SUPFAM

SSF48239. Terp_cyc_toroid. 1 hit.
SSF48576. Terpenoid_synth. 1 hit.

ProtoNet

Search...

Other

EvolutionaryTrace

Q40577.

Entry information

Entry name

5EAS_TOBAC

Accession

Primary (citable) accession number: Q40577

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 15, 1998
Last sequence update:	April 29, 2008
Last modified:	May 1, 2013
This is version 97 of the entry and version 3 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Plant Protein Annotation Program