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Q38710 (TPSDV_ABIGR) Reviewed, UniProtKB/Swiss-Prot

Last modified April 16, 2014. Version 82. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (3) | Third-party data text xml rdf/xml gff fasta
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Names and origin

Protein namesRecommended name:
Abietadiene synthase, chloroplastic
Alternative name(s):
(-)-abieta-7(8),13(14)-diene synthase
Abietadiene cyclase
Short name=AgAS
Agggabi

Including the following 2 domains:

  1. Abietadiene synthase
    EC=4.2.3.18
    Alternative name(s):
    Neoabietadiene synthase
    EC=4.2.3.132
  2. Copalyl diphosphate synthase
    EC=5.5.1.12
Gene names
Name:AS
Synonyms:ac22, ag22
OrganismAbies grandis (Grand fir) (Pinus grandis)
Taxonomic identifier46611 [NCBI]
Taxonomic lineageEukaryotaViridiplantaeStreptophytaEmbryophytaTracheophytaSpermatophytaPinidaePinalesPinaceaeAbies

Protein attributes

Sequence length868 AA.
Sequence statusComplete.
Sequence processingThe displayed sequence is further processed into a mature form.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Involved in defensive oleoresin formation in conifers in response to insect attack or other injury. Involved in diterpene (C20) olefins biosynthesis. Bifunctional enzyme that catalyzes two sequential cyclizations of geranylgeranyl diphosphate (GGPP) to abietadiene. The copalyl diphosphate (CPP) intermediate diffuses freely between the 2 active sites in the enzyme. Changes in reaction pH, but not salt concentration, influence the relative proportion of the major products of the enzyme, abitadiene, levopimaradiene and neoabitadiene. Ref.3 Ref.4 Ref.5 Ref.7 Ref.9 Ref.10 Ref.11

Catalytic activity

Geranylgeranyl diphosphate = (+)-copalyl diphosphate. Ref.4 Ref.5

+-copalyl diphosphate = abieta-7,13-diene + diphosphate. Ref.4 Ref.5

+-copalyl diphosphate = neoabietadiene + diphosphate. Ref.4 Ref.5

Cofactor

Binds 3 magnesium ions per subunit By similarity.

Potassium.

Enzyme regulation

Not inhibited by 13-cyclopropylidene or 16-methylidenegeranylgeranyl diphosphate. The copalyl diphosphate synthase activity is not susceptible to magnesium-dependent inhibition. Ref.4

Pathway

Terpene metabolism; oleoresin biosynthesis.

Subunit structure

Monomer.

Subcellular location

Plastidchloroplast By similarity.

Induction

By wounding. Ref.1 Ref.4

Domain

The Asp-Xaa-Asp-Asp (DXDD) and the Asp-Asp-Xaa-Xaa-Asp/Glu (DDXXD/E) motifs are important for the catalytic activities, presumably through binding to Mg2+.

Post-translational modification

The N-terminus is blocked.

Miscellaneous

The abietadiene synthase activity exhibits an absolute dependence on a divalent metal ion cofactor while the copalyl diphosphate synthase activity is not completely dependent.

Sequence similarities

Belongs to the terpene synthase family. Tpsd subfamily.

Biophysicochemical properties

Kinetic parameters:

KM=3 µM for GGPP (in presence of magnesium) Ref.4 Ref.7

KM=20 µM for GGPP (in absence of magnesium)

KM=0.4 µM for CPP (in presence of magnesium)

KM=10 µM for CPP (in absence of magnesium)

pH dependence:

Optimum pH is 7.2 with GGPP as substrate and 8.7 with CPP as substrate.

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Transit peptide1 – 7070Chloroplast Potential
Chain71 – 868798Abietadiene synthase, chloroplastic
PRO_0000033634

Regions

Motif402 – 4054DXDD motif
Motif621 – 6255DDXXD motif

Sites

Metal binding6211Magnesium 1 By similarity
Metal binding6211Magnesium 2 By similarity
Metal binding6251Magnesium 1 By similarity
Metal binding6251Magnesium 2 By similarity
Metal binding7651Magnesium 3 By similarity
Metal binding7691Magnesium 3 By similarity
Metal binding7731Magnesium 3 By similarity

Experimental info

Mutagenesis861K → A: No effect on CPP binding, but decreased abietadiene synthase activity; when associated with A-87. Ref.9
Mutagenesis871R → A: No effect on CPP binding, but decreased abietadiene synthase activity; when associated with A-86. Ref.9
Mutagenesis3561R → A: No effect on abietadiene synthase activity, but strongly reduced copalyl diphosphate synthase activity. Ref.11
Mutagenesis3561R → H: Increased Mg(2+) inhibition; when associated with A-621. Ref.11
Mutagenesis3581W → A: Decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis3611D → A: No effect on GGPP binding, but decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis3651R → A: No effect on GGPP binding, but decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis4021D → A, E or N: No effect on GGPP binding, but decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis4041D → A, E or N: No effect on GGPP binding, but loss of copalyl diphosphate synthase activity. Ref.6 Ref.7
Mutagenesis4051D → A, E or N: No effect on GGPP binding, but decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis4111R → A: No effect on GGPP binding, but slightly decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis4511N → A: No effect on abietadiene synthase activity, but strongly reduced copalyl diphosphate synthase activity. Ref.12
Mutagenesis4541R → A: No effect on GGPP binding, but decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis4991E → A: No effect on GGPP binding, but decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis5201Y → A: No effect on GGPP binding, but slightly decreased copalyl diphosphate synthase activity. Ref.7
Mutagenesis5841R → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis5861R → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis5891E → A: Lower substrate binding and strong decrease of abietadiene synthase activity. Ref.8
Mutagenesis6171T → A: Increased production of abietadiene at the expense of levopimaradiene. Ref.8
Mutagenesis6211D → A: Loss of abietadiene synthase activity. Increased Mg(2+) inhibition; when associated with H-356. Ref.6 Ref.8 Ref.11
Mutagenesis6251D → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis6991E → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis7211S → A: Lower substrate binding and strong decrease of abietadiene synthase activity. Ref.8
Mutagenesis7231A → S: Produces pimaradienes instead of abietadienes. Ref.10
Mutagenesis7271V → T: No effect. Ref.12
Mutagenesis7621R → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis7651N → A: Abolishes the conversion of CPP to abietadiene; produces only sandaracopimaradiene. Ref.8
Mutagenesis7661D → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis7691T → A: Increased production of neoabietadiene at the expense of both levopimaradiene and abietadiene. Ref.8
Mutagenesis7731E → A: Increased production of neoabietadiene. Ref.8
Mutagenesis7781E → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis8411Y → F: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis8451D → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Mutagenesis8481T → A: No or small effect on substrate binding, but strongly decreases abietadiene synthase activity. Ref.8
Sequence conflict231A → T in AAK83563. Ref.2
Sequence conflict2141F → S in AAK83563. Ref.2
Sequence conflict297 – 2993DWQ → EWE in AAK83563. Ref.2
Sequence conflict5571V → L in AAK83563. Ref.2
Sequence conflict5791D → E in AAK83563. Ref.2
Sequence conflict6531Q → K in AAK83563. Ref.2

Secondary structure

............................................................................................... 868
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
Q38710 [UniParc].

Last modified November 1, 1996. Version 1.
Checksum: AD5E79F56B70D25C

FASTA86899,536
        10         20         30         40         50         60 
MAMPSSSLSS QIPTAAHHLT ANAQSIPHFS TTLNAGSSAS KRRSLYLRWG KGSNKIIACV 

        70         80         90        100        110        120 
GEGGATSVPY QSAEKNDSLS SSTLVKREFP PGFWKDDLID SLTSSHKVAA SDEKRIETLI 

       130        140        150        160        170        180 
SEIKNMFRCM GYGETNPSAY DTAWVARIPA VDGSDNPHFP ETVEWILQNQ LKDGSWGEGF 

       190        200        210        220        230        240 
YFLAYDRILA TLACIITLTL WRTGETQVQK GIEFFRTQAG KMEDEADSHR PSGFEIVFPA 

       250        260        270        280        290        300 
MLKEAKILGL DLPYDLPFLK QIIEKREAKL KRIPTDVLYA LPTTLLYSLE GLQEIVDWQK 

       310        320        330        340        350        360 
IMKLQSKDGS FLSSPASTAA VFMRTGNKKC LDFLNFVLKK FGNHVPCHYP LDLFERLWAV 

       370        380        390        400        410        420 
DTVERLGIDR HFKEEIKEAL DYVYSHWDER GIGWARENPV PDIDDTAMGL RILRLHGYNV 

       430        440        450        460        470        480 
SSDVLKTFRD ENGEFFCFLG QTQRGVTDML NVNRCSHVSF PGETIMEEAK LCTERYLRNA 

       490        500        510        520        530        540 
LENVDAFDKW AFKKNIRGEV EYALKYPWHK SMPRLEARSY IENYGPDDVW LGKTVYMMPY 

       550        560        570        580        590        600 
ISNEKYLELA KLDFNKVQSI HQTELQDLRR WWKSSGFTDL NFTRERVTEI YFSPASFIFE 

       610        620        630        640        650        660 
PEFSKCREVY TKTSNFTVIL DDLYDAHGSL DDLKLFTESV KRWDLSLVDQ MPQQMKICFV 

       670        680        690        700        710        720 
GFYNTFNDIA KEGRERQGRD VLGYIQNVWK VQLEAYTKEA EWSEAKYVPS FNEYIENASV 

       730        740        750        760        770        780 
SIALGTVVLI SALFTGEVLT DEVLSKIDRE SRFLQLMGLT GRLVNDTKTY QAERGQGEVA 

       790        800        810        820        830        840 
SAIQCYMKDH PKISEEEALQ HVYSVMENAL EELNREFVNN KIPDIYKRLV FETARIMQLF 

       850        860 
YMQGDGLTLS HDMEIKEHVK NCLFQPVA 

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References

[1]"Abietadiene synthase from grand fir (Abies grandis). cDNA isolation, characterization, and bacterial expression of a bifunctional diterpene cyclase involved in resin acid biosynthesis."
Stoffer Vogel B., Wildung M.R., Vogel G., Croteau R.B.
J. Biol. Chem. 271:23262-23268(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 148-166; 522-530; 615-625 AND 699-705, INDUCTION.
Tissue: Stem.
[2]"Genomic organization of plant terpene synthases and molecular evolutionary implications."
Trapp S.C., Croteau R.B.
Genetics 158:811-832(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 16-868, NOMENCLATURE.
[3]"Plant terpenoid synthases: molecular biology and phylogenetic analysis."
Bohlmann J., Meyer-Gauen G., Croteau R.B.
Proc. Natl. Acad. Sci. U.S.A. 95:4126-4133(1998) [PubMed] [Europe PMC] [Abstract]
Cited for: GENE FAMILY, NOMENCLATURE, FUNCTION.
[4]"Abietadiene synthase from grand fir (Abies grandis): characterization and mechanism of action of the 'pseudomature' recombinant enzyme."
Peters R.J., Flory J.E., Jetter R., Ravn M.M., Lee H.J., Coates R.M., Croteau R.B.
Biochemistry 39:15592-15602(2000) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION.
[5]"Stereochemistry of the cyclization-rearrangement of (+)-copalyl diphosphate to (-)-abietadiene catalyzed by recombinant abietadiene synthase from Abies grandis."
Ravn M.M., Coates R.M., Flory J.E., Peters R.J., Croteau R.
Org. Lett. 2:573-576(2000) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, CATALYTIC ACTIVITY.
[6]"Bifunctional abietadiene synthase: free diffusive transfer of the (+)-copalyl diphosphate intermediate between two distinct active sites."
Peters R.J., Ravn M.M., Coates R.M., Croteau R.B.
J. Am. Chem. Soc. 123:8974-8978(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: MUTAGENESIS OF ASP-404 AND ASP-621, CHARACTERIZATION.
[7]"Abietadiene synthase catalysis: conserved residues involved in protonation-initiated cyclization of geranylgeranyl diphosphate to (+)-copalyl diphosphate."
Peters R.J., Croteau R.B.
Biochemistry 41:1836-1842(2002) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, 3D-STRUCTURE MODELING, MUTAGENESIS OF TRP-358; ASP-361; ARG-365; ASP-402; ASP-404; ASP-405; ARG-411; ARG-454; GLU-499 AND TYR-520.
[8]"Abietadiene synthase catalysis: mutational analysis of a prenyl diphosphate ionization-initiated cyclization and rearrangement."
Peters R.J., Croteau R.B.
Proc. Natl. Acad. Sci. U.S.A. 99:580-584(2002) [PubMed] [Europe PMC] [Abstract]
Cited for: MUTAGENESIS OF ARG-584; ARG-586; GLU-589; THR-617; ASP-621; ASP-625; GLU-699; SER-721; ARG-762; ASN-765; ASP-766; THR-769; GLU-773; GLU-778; TYR-841; ASP-845 AND THR-848, CHARACTERIZATION.
[9]"Bifunctional abietadiene synthase: mutual structural dependence of the active sites for protonation-initiated and ionization-initiated cyclizations."
Peters R.J., Carter O.A., Zhang Y., Matthews B.W., Croteau R.B.
Biochemistry 42:2700-2707(2003) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, MUTAGENESIS OF LYS-86 AND ARG-87.
[10]"A single residue switch converts abietadiene synthase into a pimaradiene specific cyclase."
Wilderman P.R., Peters R.J.
J. Am. Chem. Soc. 129:15736-15737(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, MUTAGENESIS OF ALA-723.
[11]"A single residue switch for Mg(2+)-dependent inhibition characterizes plant class II diterpene cyclases from primary and secondary metabolism."
Mann F.M., Prisic S., Davenport E.K., Determan M.K., Coates R.M., Peters R.J.
J. Biol. Chem. 285:20558-20563(2010) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, MUTAGENESIS OF ARG-356 AND ASP-621.
[12]"Insights into diterpene cyclization from structure of bifunctional abietadiene synthase from Abies grandis."
Zhou K., Gao Y., Hoy J.A., Mann F.M., Honzatko R.B., Peters R.J.
J. Biol. Chem. 287:6840-6850(2012) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) OF 85-868, MUTAGENESIS OF ASN-451 AND VAL-727.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
U50768 mRNA. Translation: AAB05407.1.
AF326516 Genomic DNA. Translation: AAK83563.1.

3D structure databases

PDBe
RCSB-PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
3S9VX-ray2.30A/B/C/D85-868[»]
ProteinModelPortalQ38710.
ModBaseSearch...
MobiDBSearch...

Proteomic databases

PRIDEQ38710.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Enzyme and pathway databases

BioCycMetaCyc:MONOMER-12822.
BRENDA4.2.3.18. 2.
UniPathwayUPA00924.

Family and domain databases

Gene3D1.10.600.10. 1 hit.
1.50.10.20. 1 hit.
1.50.30.10. 1 hit.
InterProIPR001906. Terpene_synth_N.
IPR005630. Terpene_synthase_metal-bd.
IPR008930. Terpenoid_cyclase/PrenylTrfase.
IPR008949. Terpenoid_synth.
[Graphical view]
PfamPF01397. Terpene_synth. 1 hit.
PF03936. Terpene_synth_C. 1 hit.
[Graphical view]
SUPFAMSSF48239. SSF48239. 3 hits.
SSF48576. SSF48576. 1 hit.
ProtoNetSearch...

Entry information

Entry nameTPSDV_ABIGR
AccessionPrimary (citable) accession number: Q38710
Secondary accession number(s): Q94FW1
Entry history
Integrated into UniProtKB/Swiss-Prot: April 12, 2005
Last sequence update: November 1, 1996
Last modified: April 16, 2014
This is version 82 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programPlant Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

PATHWAY comments

Index of metabolic and biosynthesis pathways