PIKA4_STRVZ
ID PIKA4_STRVZ Reviewed; 1346 AA.
AC Q9ZGI2;
DT 08-JUN-2016, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-1999, sequence version 1.
DT 03-AUG-2022, entry version 124.
DE RecName: Full=Narbonolide/10-deoxymethynolide synthase PikA4, module 6 {ECO:0000305};
DE EC=2.3.1.239 {ECO:0000269|PubMed:10421766, ECO:0000305|PubMed:12379101, ECO:0000305|PubMed:12733905, ECO:0000305|PubMed:17719493};
DE EC=2.3.1.240 {ECO:0000269|PubMed:10421766, ECO:0000305|PubMed:12379101, ECO:0000305|PubMed:12733905, ECO:0000305|PubMed:17719493};
DE AltName: Full=Narbonolide/10-deoxymethynolide synthase PikAIV {ECO:0000305};
DE AltName: Full=Pikromycin polyketide synthase component PikAIV {ECO:0000303|PubMed:10421766};
DE Short=Pikromycin PKS component PikAIV {ECO:0000303|PubMed:10421766};
DE AltName: Full=Type I modular polyketide synthase PikAIV {ECO:0000303|PubMed:10421766};
DE Short=PKS {ECO:0000303|PubMed:10421766};
GN Name=pikAIV {ECO:0000303|PubMed:9770448};
OS Streptomyces venezuelae.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=54571;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PATHWAY.
RC STRAIN=ATCC 15439 / DSM 41110 / IMRU3627 / M-2140;
RX PubMed=9770448; DOI=10.1073/pnas.95.21.12111;
RA Xue Y., Zhao L., Liu H.W., Sherman D.H.;
RT "A gene cluster for macrolide antibiotic biosynthesis in Streptomyces
RT venezuelae: architecture of metabolic diversity.";
RL Proc. Natl. Acad. Sci. U.S.A. 95:12111-12116(1998).
RN [2]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=10421766; DOI=10.1016/s1074-5521(99)80087-8;
RA Tang L., Fu H., Betlach M.C., McDaniel R.;
RT "Elucidating the mechanism of chain termination switching in the
RT picromycin/methymycin polyketide synthase.";
RL Chem. Biol. 6:553-558(1999).
RN [3]
RP FUNCTION, AND DOMAIN.
RC STRAIN=ATCC 15439 / DSM 41110 / IMRU3627 / M-2140;
RX PubMed=10676969; DOI=10.1038/35000624;
RA Xue Y., Sherman D.H.;
RT "Alternative modular polyketide synthase expression controls macrolactone
RT structure.";
RL Nature 403:571-575(2000).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF
RP SER-1196; ASP-1224; GLU-1235 AND ARG-1239, AND SUBSTRATE SPECIFICITY.
RX PubMed=12379101; DOI=10.1021/bi026006d;
RA Lu H., Tsai S.C., Khosla C., Cane D.E.;
RT "Expression, site-directed mutagenesis, and steady state kinetic analysis
RT of the terminal thioesterase domain of the methymycin/picromycin polyketide
RT synthase.";
RL Biochemistry 41:12590-12597(2002).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, AND SUBSTRATE SPECIFICITY.
RX PubMed=12733905; DOI=10.1021/ja034574q;
RA Yin Y., Lu H., Khosla C., Cane D.E.;
RT "Expression and kinetic analysis of the substrate specificity of modules 5
RT and 6 of the picromycin/methymycin polyketide synthase.";
RL J. Am. Chem. Soc. 125:5671-5676(2003).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF CYS-207; SER-652; SER-980 AND
RP SER-1196, COFACTOR, ACTIVE SITE, AND PHOSPHOPANTETHEINYLATION AT SER-980.
RX PubMed=17719493; DOI=10.1016/j.chembiol.2007.07.013;
RA Kittendorf J.D., Beck B.J., Buchholz T.J., Seufert W., Sherman D.H.;
RT "Interrogating the molecular basis for multiple macrolactone ring formation
RT by the pikromycin polyketide synthase.";
RL Chem. Biol. 14:944-954(2007).
RN [7]
RP FUNCTION, PATHWAY, AND SUBUNIT.
RX PubMed=19027305; DOI=10.1016/j.bmc.2008.10.082;
RA Kittendorf J.D., Sherman D.H.;
RT "The methymycin/pikromycin pathway: a model for metabolic diversity in
RT natural product biosynthesis.";
RL Bioorg. Med. Chem. 17:2137-2146(2009).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF 1049-1346, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBUNIT.
RX PubMed=12379102; DOI=10.1021/bi0260177;
RA Tsai S.-C., Lu H., Cane D.E., Khosla C., Stroud R.M.;
RT "Insights into channel architecture and substrate specificity from crystal
RT structures of two macrocycle-forming thioesterases of modular polyketide
RT synthases.";
RL Biochemistry 41:12598-12606(2002).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) OF 1049-1346 IN COMPLEX WITH
RP SUBSTRATE ANALOGS, ACTIVITY REGULATION, ACTIVE SITE, AND SUBUNIT.
RX PubMed=16969373; DOI=10.1038/nchembio822;
RA Giraldes J.W., Akey D.L., Kittendorf J.D., Sherman D.H., Smith J.L.,
RA Fecik R.A.;
RT "Structural and mechanistic insights into polyketide macrolactonization
RT from polyketide-based affinity labels.";
RL Nat. Chem. Biol. 2:531-536(2006).
RN [10]
RP X-RAY CRYSTALLOGRAPHY (1.79 ANGSTROMS) OF 1049-1346 OF WILD-TYPE AND MUTANT
RP ALA-1196 IN COMPLEX WITH SUBSTRATE ANALOGS, FUNCTION, MUTAGENESIS OF
RP SER-1196, REACTION MECHANISM, ACTIVE SITE, AND SUBUNIT.
RX PubMed=16969372; DOI=10.1038/nchembio824;
RA Akey D.L., Kittendorf J.D., Giraldes J.W., Fecik R.A., Sherman D.H.,
RA Smith J.L.;
RT "Structural basis for macrolactonization by the pikromycin thioesterase.";
RL Nat. Chem. Biol. 2:537-542(2006).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) OF 1-37, AND SUBUNIT.
RX PubMed=19146481; DOI=10.1021/cb8002607;
RA Buchholz T.J., Geders T.W., Bartley F.E., Reynolds K.A., Smith J.L.,
RA Sherman D.H.;
RT "Structural basis for binding specificity between subclasses of modular
RT polyketide synthase docking domains.";
RL ACS Chem. Biol. 4:41-52(2009).
CC -!- FUNCTION: Involved in the biosynthesis of 12- and 14-membered ring
CC macrolactone antibiotics such as methymycin and neomethymycin, and
CC pikromycin and narbomycin, respectively. Component of the pikromycin
CC PKS which catalyzes the biosynthesis of both precursors 10-
CC deoxymethynolide (12-membered ring macrolactone) and narbonolide (14-
CC membered ring macrolactone). Chain elongation through PikAI, PikAII and
CC PikAIII followed by thioesterase catalyzed termination results in the
CC production of 10-deoxymethynolide, while continued elongation through
CC PikAIV, followed by thioesterase (TE) catalyzed cyclization results in
CC the biosynthesis of the narbonolide. The thioesterase can use a series
CC of diketide-N-acetylcysteamine (SNAC) thioesters, but has a strong
CC preference for the 2-methyl-3-ketopentanoyl-SNAC over the stereoisomers
CC of 2-methyl-3-hydroxyacyl-SNAC (PubMed:12379101, PubMed:12733905).
CC {ECO:0000269|PubMed:10421766, ECO:0000269|PubMed:10676969,
CC ECO:0000269|PubMed:12379101, ECO:0000269|PubMed:12733905,
CC ECO:0000269|PubMed:16969372, ECO:0000269|PubMed:17719493,
CC ECO:0000305|PubMed:19027305}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=5 (S)-methylmalonyl-CoA + 11 H(+) + malonyl-CoA + 5 NADPH =
CC 10-deoxymethynolide + 6 CO2 + 6 CoA + 2 H2O + 5 NADP(+);
CC Xref=Rhea:RHEA:43056, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16526, ChEBI:CHEBI:29461, ChEBI:CHEBI:57287,
CC ChEBI:CHEBI:57327, ChEBI:CHEBI:57384, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; EC=2.3.1.239;
CC Evidence={ECO:0000269|PubMed:10421766, ECO:0000305|PubMed:12379101,
CC ECO:0000305|PubMed:12733905, ECO:0000305|PubMed:17719493};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=6 (S)-methylmalonyl-CoA + 12 H(+) + malonyl-CoA + 5 NADPH = 7
CC CO2 + 7 CoA + 2 H2O + 5 NADP(+) + narbonolide; Xref=Rhea:RHEA:42844,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:16526,
CC ChEBI:CHEBI:29650, ChEBI:CHEBI:57287, ChEBI:CHEBI:57327,
CC ChEBI:CHEBI:57384, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC EC=2.3.1.240; Evidence={ECO:0000269|PubMed:10421766,
CC ECO:0000305|PubMed:12379101, ECO:0000305|PubMed:12733905,
CC ECO:0000305|PubMed:17719493};
CC -!- COFACTOR:
CC Name=pantetheine 4'-phosphate; Xref=ChEBI:CHEBI:47942;
CC Evidence={ECO:0000305|PubMed:17719493};
CC Note=Binds 1 phosphopantetheine covalently. {ECO:0000305};
CC -!- ACTIVITY REGULATION: Irreversibly inhibited by (2S,3R,4S)-2,4-
CC dihydroxy-3-methylhexyl-phosphonic acid and (3R,4S)-4-hydroxy-3-methyl-
CC 2-oxohexyl-phosphonic acid. {ECO:0000269|PubMed:16969373}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=7.3 mM for (2R,3S)-2-methyl-3-hydroxyacyl-SNAC (thioesterase
CC activity) {ECO:0000269|PubMed:12379101};
CC KM=13 mM for (2S,3S)-2-methyl-3-hydroxyacyl-SNAC (thioesterase
CC activity) {ECO:0000269|PubMed:12379101};
CC KM=15 mM for 2-methyl-3-ketopentanoyl-SNAC (thioesterase activity)
CC {ECO:0000269|PubMed:12379101};
CC KM=16 mM for (2R,3R)-2-methyl-3-hydroxyacyl-SNAC (thioesterase
CC activity) {ECO:0000269|PubMed:12379101};
CC KM=21 mM for (2S,3R)-2-methyl-3-hydroxyacyl-SNAC (thioesterase
CC activity) {ECO:0000269|PubMed:12379101};
CC Note=kcat is 56 min(-1) for thioesterase activity with 2-methyl-3-
CC ketopentanoyl-SNAC as substrate. kcat is 7.9 min(-1) for thioesterase
CC activity with (2S,3R)-2-methyl-3-hydroxyacyl-SNAC as substrate. kcat
CC is 4.3 min(-1) for thioesterase activity with (2R,3S)-2-methyl-3-
CC hydroxyacyl-SNAC as substrate. kcat is 2.6 min(-1) for thioesterase
CC activity with (2S,3S)-2-methyl-3-hydroxyacyl-SNAC as substrate. kcat
CC is 1.8 min(-1) for thioesterase activity with (2R,3R)-2-methyl-3-
CC hydroxyacyl-SNAC as substrate. {ECO:0000269|PubMed:12379101};
CC pH dependence:
CC Optimum pH is between 8 and 8.4 (PubMed:12379101). In the
CC thioesterase domain, the size of the substrate channel increases with
CC increasing pH (PubMed:12379102). {ECO:0000269|PubMed:12379101,
CC ECO:0000269|PubMed:12379102};
CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000305|PubMed:19027305,
CC ECO:0000305|PubMed:9770448}.
CC -!- SUBUNIT: Homodimer (PubMed:12379102, PubMed:16969373, PubMed:16969372,
CC PubMed:19146481). Pikromycin PKS consists of a combination of
CC multimodular (PikAI and PikAII) and monomodular (PikAIII and PikAIV)
CC polypeptides each coding for a functional synthase subunit which
CC participates in 1 (monomodular) or 2 (multimodular) of the six FAS-like
CC elongation steps required for formation of the polyketide. Module 1, 2,
CC 3, 4, 5, and 6 participating in biosynthesis steps 1, 2, 3, 4, 5, and
CC 6, respectively. {ECO:0000269|PubMed:12379102,
CC ECO:0000269|PubMed:16969372, ECO:0000269|PubMed:16969373,
CC ECO:0000269|PubMed:19146481, ECO:0000305|PubMed:19027305}.
CC -!- DOMAIN: Cells lacking the TE domain are unable to produce methymycin,
CC neomethymycin, narbomycin and pikromycin.
CC {ECO:0000269|PubMed:10676969}.
CC -!- MISCELLANEOUS: Type I modular polyketide synthases (PKSs) catalyze the
CC step-wise condensation of simple carboxylic acid derivatives.
CC Organizationally, type I PKSs are arranged into modules, wherein each
CC module is comprised of a set of catalytic activities that is
CC responsible for a single elongation of the polyketide chain and the
CC appropriate reductive processing of the beta-keto functionality. A
CC minimal elongation module contains an acyl transferase (AT) domain, an
CC acyl-carrier protein (ACP) domain, and a ketosynthase (KS) domain. The
CC AT domain is responsible for loading the methylmalonyl-CoA extender
CC unit onto the phosphopantetheinylated ACP domain. Subsequently, the KS
CC domain decarboxylates and then condenses the ACP-bound extender unit
CC with the growing polyketide chain obtained from the preceding module to
CC yield an ACP-bound beta-ketoacyl intermediate. In addition to the three
CC core domains, each elongation module may contain up to three additional
CC domains: a ketoreductase (KR), dehydratase (DH), and an enoyl reductase
CC (ER) that are responsible for the reductive processing of the beta-keto
CC functionality prior to the next extension step. The presence of a KR
CC domain alone gives rise to a beta-hydroxyl functionality, the presence
CC of both a KR and a DH domain generates an alkene, while the combination
CC of KR, DH, and ER results in complete reduction to the alkane. Finally,
CC a thioesterase (TE) domain, typically found at the terminus of the last
CC elongation module, catalyzes the termination of polyketide
CC biosynthesis. The activity of this domain results in cleavage of the
CC acyl chain from the adjacent ACP and formation of the macrocyclic ring.
CC {ECO:0000305|PubMed:19027305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; AF079138; AAC69332.1; -; Genomic_DNA.
DR PIR; T17412; T17412.
DR PDB; 1MN6; X-ray; 2.20 A; A/B=1049-1346.
DR PDB; 1MNA; X-ray; 1.80 A; A/B=1049-1346.
DR PDB; 1MNQ; X-ray; 2.20 A; A/B=1049-1346.
DR PDB; 2H7X; X-ray; 1.85 A; A/B=1049-1346.
DR PDB; 2H7Y; X-ray; 2.10 A; A/B=1049-1346.
DR PDB; 2HFJ; X-ray; 1.95 A; A/B=1049-1346.
DR PDB; 2HFK; X-ray; 1.79 A; A/B=1049-1346.
DR PDB; 3F5H; X-ray; 1.75 A; A/B=1-37.
DR PDBsum; 1MN6; -.
DR PDBsum; 1MNA; -.
DR PDBsum; 1MNQ; -.
DR PDBsum; 2H7X; -.
DR PDBsum; 2H7Y; -.
DR PDBsum; 2HFJ; -.
DR PDBsum; 2HFK; -.
DR PDBsum; 3F5H; -.
DR AlphaFoldDB; Q9ZGI2; -.
DR SMR; Q9ZGI2; -.
DR DrugBank; DB07703; (3R,4S,5S,7R,9E,11R,12R)-12-ETHYL-4-HYDROXY-3,5,7,11-TETRAMETHYLOXACYCLODODEC-9-ENE-2,8-DIONE.
DR DrugBank; DB08431; [(3R,4S)-4-HYDROXY-3-METHYL-2-OXOHEXYL]PHOSPHONIC ACID.
DR DrugBank; DB08759; [(3R,4S,5S,7R)-4,8-DIHYDROXY-3,5,7-TRIMETHYL-2-OXOOCTYL]PHOSPHONIC ACID.
DR ESTHER; strve-PIKAIV; Thioesterase.
DR KEGG; ag:AAC69332; -.
DR BioCyc; MetaCyc:MON-18414; -.
DR BRENDA; 2.3.1.239; 6106.
DR BRENDA; 2.3.1.240; 6106.
DR EvolutionaryTrace; Q9ZGI2; -.
DR GO; GO:0004315; F:3-oxoacyl-[acyl-carrier-protein] synthase activity; IEA:InterPro.
DR GO; GO:0016747; F:acyltransferase activity, transferring groups other than amino-acyl groups; IDA:UniProtKB.
DR GO; GO:0031177; F:phosphopantetheine binding; TAS:UniProtKB.
DR GO; GO:0006633; P:fatty acid biosynthetic process; IEA:InterPro.
DR GO; GO:0033068; P:macrolide biosynthetic process; IDA:UniProtKB.
DR Gene3D; 1.10.1200.10; -; 1.
DR Gene3D; 3.40.366.10; -; 1.
DR Gene3D; 3.40.47.10; -; 1.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR001227; Ac_transferase_dom_sf.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR014043; Acyl_transferase.
DR InterPro; IPR016035; Acyl_Trfase/lysoPLipase.
DR InterPro; IPR018201; Ketoacyl_synth_AS.
DR InterPro; IPR014031; Ketoacyl_synth_C.
DR InterPro; IPR014030; Ketoacyl_synth_N.
DR InterPro; IPR016036; Malonyl_transacylase_ACP-bd.
DR InterPro; IPR032821; PKS_assoc.
DR InterPro; IPR020841; PKS_Beta-ketoAc_synthase_dom.
DR InterPro; IPR020806; PKS_PP-bd.
DR InterPro; IPR020802; PKS_thioesterase.
DR InterPro; IPR015083; Polyketide_synth_docking.
DR InterPro; IPR009081; PP-bd_ACP.
DR InterPro; IPR001031; Thioesterase.
DR InterPro; IPR016039; Thiolase-like.
DR Pfam; PF00698; Acyl_transf_1; 1.
DR Pfam; PF08990; Docking; 1.
DR Pfam; PF16197; KAsynt_C_assoc; 1.
DR Pfam; PF00109; ketoacyl-synt; 1.
DR Pfam; PF02801; Ketoacyl-synt_C; 1.
DR Pfam; PF00550; PP-binding; 1.
DR Pfam; PF00975; Thioesterase; 1.
DR SMART; SM00827; PKS_AT; 1.
DR SMART; SM00825; PKS_KS; 1.
DR SMART; SM00823; PKS_PP; 1.
DR SMART; SM00824; PKS_TE; 1.
DR SUPFAM; SSF47336; SSF47336; 1.
DR SUPFAM; SSF52151; SSF52151; 1.
DR SUPFAM; SSF53474; SSF53474; 1.
DR SUPFAM; SSF53901; SSF53901; 1.
DR SUPFAM; SSF55048; SSF55048; 1.
DR PROSITE; PS00606; B_KETOACYL_SYNTHASE; 1.
DR PROSITE; PS50075; CARRIER; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acyltransferase; Antibiotic biosynthesis; Coiled coil;
KW Multifunctional enzyme; NADP; Phosphopantetheine; Phosphoprotein;
KW Transferase.
FT CHAIN 1..1346
FT /note="Narbonolide/10-deoxymethynolide synthase PikA4,
FT module 6"
FT /id="PRO_0000436360"
FT DOMAIN 945..1020
FT /note="Carrier"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT REGION 37..1332
FT /note="Module 6"
FT /evidence="ECO:0000305"
FT REGION 37..464
FT /note="Beta-ketoacyl synthase"
FT /evidence="ECO:0000305"
FT REGION 562..844
FT /note="Acyltransferase"
FT /evidence="ECO:0000305"
FT REGION 1028..1050
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 1127..1332
FT /note="Thioesterase"
FT /evidence="ECO:0000305"
FT COILED 3..32
FT /evidence="ECO:0000255"
FT ACT_SITE 207
FT /note="For beta-ketoacyl synthase activity"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10022"
FT ACT_SITE 652
FT /note="Acyl-ester intermediate; for acyltransferase
FT activity"
FT /evidence="ECO:0000250|UniProtKB:Q03133,
FT ECO:0000305|PubMed:17719493"
FT ACT_SITE 1196
FT /note="Nucleophile; for thioesterase activity"
FT /evidence="ECO:0000250|UniProtKB:Q03133,
FT ECO:0000269|PubMed:16969372, ECO:0000305|PubMed:16969373"
FT ACT_SITE 1316
FT /note="Proton acceptor; for thioesterase activity"
FT /evidence="ECO:0000250|UniProtKB:Q03133,
FT ECO:0000305|PubMed:16969372, ECO:0000305|PubMed:16969373"
FT BINDING 1125
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:16969372"
FT BINDING 1197
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:16969372,
FT ECO:0000269|PubMed:16969373"
FT BINDING 1224
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q03133"
FT MOD_RES 980
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258,
FT ECO:0000305|PubMed:17719493"
FT MUTAGEN 207
FT /note="C->A: Unable to catalyze the biosynthesis of the 14-
FT membered ring macrolactone narbonolide, however it is able
FT to produce the 12-membered ring macrolactone 10-
FT deoxymethynolide."
FT /evidence="ECO:0000269|PubMed:17719493"
FT MUTAGEN 652
FT /note="S->A: Unable to catalyze the biosynthesis of the 14-
FT membered ring macrolactone narbonolide, however it is able
FT to produce the 12-membered ring macrolactone 10-
FT deoxymethynolide."
FT /evidence="ECO:0000269|PubMed:17719493"
FT MUTAGEN 980
FT /note="S->A: Unable to catalyze the biosynthesis of the 14-
FT membered ring macrolactone narbonolide, however it is able
FT to produce the 12-membered ring macrolactone 10-
FT deoxymethynolide."
FT /evidence="ECO:0000269|PubMed:17719493"
FT MUTAGEN 1196
FT /note="S->A: Loss of thioesterase activity. Unable to
FT catalyze the biosynthesis of the 14-membered ring
FT macrolactone narbonolide and 12-membered ring macrolactone
FT 10-deoxymethynolide. The 10-deoxymethynolide molecule is
FT removed from the catalytic triad, does not reach the most
FT hydrophobic region of the channel, disrupts the
FT hydrophilic-barrier water network and is rotated with
FT respect the phosphopentaketide."
FT /evidence="ECO:0000269|PubMed:12379101,
FT ECO:0000269|PubMed:16969372, ECO:0000269|PubMed:17719493"
FT MUTAGEN 1224
FT /note="D->A: Retains significant albeit reduced
FT thioesterase activity."
FT /evidence="ECO:0000269|PubMed:12379101"
FT MUTAGEN 1235
FT /note="E->N: Only relatively minor changes in the
FT thioesterase activity. 10-fold reduction in the kcat/Km for
FT ketoester; when associated with E-1239."
FT /evidence="ECO:0000269|PubMed:12379101"
FT MUTAGEN 1239
FT /note="R->E: 10-fold reduction in the kcat/Km for
FT ketoester; when associated with N-1235."
FT /evidence="ECO:0000269|PubMed:12379101"
FT MUTAGEN 1239
FT /note="R->N: 3-fold decrease fo the catalytic efficiency
FT and 3-fold increase of affinity."
FT /evidence="ECO:0000269|PubMed:12379101"
FT HELIX 1..32
FT /evidence="ECO:0007829|PDB:3F5H"
FT HELIX 1059..1069
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1073..1084
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1089..1092
FT /evidence="ECO:0007829|PDB:1MNA"
FT HELIX 1093..1095
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1103..1106
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1118..1122
FT /evidence="ECO:0007829|PDB:2HFK"
FT TURN 1131..1134
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1135..1139
FT /evidence="ECO:0007829|PDB:2HFK"
FT TURN 1140..1144
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1147..1150
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1166..1169
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1170..1185
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1190..1195
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1197..1213
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1218..1224
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1232..1236
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1238..1247
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1255..1269
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1280..1287
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1294..1296
FT /evidence="ECO:0007829|PDB:2HFK"
FT STRAND 1307..1314
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1318..1321
FT /evidence="ECO:0007829|PDB:2HFK"
FT HELIX 1324..1339
FT /evidence="ECO:0007829|PDB:2HFK"
SQ SEQUENCE 1346 AA; 141914 MW; 3E149C8044FBE5F2 CRC64;
MTSSNEQLVD ALRASLKENE ELRKESRRRA DRRQEPMAIV GMSCRFAGGI RSPEDLWDAV
AAGKDLVSEV PEERGWDIDS LYDPVPGRKG TTYVRNAAFL DDAAGFDAAF FGISPREALA
MDPQQRQLLE ASWEVFERAG IDPASVRGTD VGVYVGCGYQ DYAPDIRVAP EGTGGYVVTG
NSSAVASGRI AYSLGLEGPA VTVDTACSSS LVALHLALKG LRNGDCSTAL VGGVAVLATP
GAFIEFSSQQ AMAADGRTKG FASAADGLAW GEGVAVLLLE RLSDARRKGH RVLAVVRGSA
INQDGASNGL TAPHGPSQQH LIRQALADAR LTSSDVDVVE GHGTGTRLGD PIEAQALLAT
YGQGRAPGQP LRLGTLKSNI GHTQAASGVA GVIKMVQALR HGVLPKTLHV DEPTDQVDWS
AGSVELLTEA VDWPERPGRL RRAGVSAFGV GGTNAHVVLE EAPAVEESPA VEPPAGGGVV
PWPVSAKTSA ALDAQIGQLA AYAEDRTDVD PAVAARALVD SRTAMEHRAV AVGDSREALR
DALRMPEGLV RGTVTDPGRV AFVFPGQGTQ WAGMGAELLD SSPEFAAAMA ECETALSPYV
DWSLEAVVRQ APSAPTLDRV DVVQPVTFAV MVSLAKVWQH HGITPEAVIG HSQGEIAAAY
VAGALTLDDA ARVVTLRSKS IAAHLAGKGG MISLALSEEA TRQRIENLHG LSIAAVNGPT
ATVVSGDPTQ IQELAQACEA DGIRARIIPV DYASHSAHVE TIENELADVL AGLSPQTPQV
PFFSTLEGTW ITEPALDGGY WYRNLRHRVG FAPAVETLAT DEGFTHFIEV SAHPVLTMTL
PDKVTGLATL RREDGGQHRL TTSLAEAWAN GLALDWASLL PATGALSPAV PDLPTYAFQH
RSYWISPAGP GEAPAHTASG REAVAETGLA WGPGAEDLDE EGRRSAVLAM VMRQAASVLR
CDSPEEVPVD RPLREIGFDS LTAVDFRNRV NRLTGLQLPP TVVFQHPTPV ALAERISDEL
AERNWAVAEP SDHEQAEEEK AAAPAGARSG ADTGAGAGMF RALFRQAVED DRYGEFLDVL
AEASAFRPQF ASPEACSERL DPVLLAGGPT DRAEGRAVLV GCTGTAANGG PHEFLRLSTS
FQEERDFLAV PLPGYGTGTG TGTALLPADL DTALDAQARA ILRAAGDAPV VLLGHSGGAL
LAHELAFRLE RAHGAPPAGI VLVDPYPPGH QEPIEVWSRQ LGEGLFAGEL EPMSDARLLA
MGRYARFLAG PRPGRSSAPV LLVRASEPLG DWQEERGDWR AHWDLPHTVA DVPGDHFTMM
RDHAPAVAEA VLSWLDAIEG IEGAGK
