ID   MALG_MALAU              Reviewed;        2345 AA.
AC   L0E2Z1;
DT   11-DEC-2019, integrated into UniProtKB/Swiss-Prot.
DT   06-MAR-2013, sequence version 1.
DT   03-AUG-2022, entry version 42.
DE   RecName: Full=Nonribisomal peptide synthase malG {ECO:0000303|PubMed:23213353};
DE            Short=NRPS malG {ECO:0000303|PubMed:23213353};
DE            EC=1.-.-.- {ECO:0000269|PubMed:31548667};
DE            EC=6.3.1.- {ECO:0000269|PubMed:31548667};
DE   AltName: Full=Malbrancheamide biosynthesis cluster protein G {ECO:0000303|PubMed:23213353};
GN   Name=malG {ECO:0000303|PubMed:23213353};
OS   Malbranchea aurantiaca.
OC   Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC   Eurotiomycetidae; Onygenales; Onygenaceae; Malbranchea.
OX   NCBI_TaxID=78605;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND DOMAIN.
RC   STRAIN=RRC1813;
RX   PubMed=23213353; DOI=10.1039/c2md20029e;
RA   Li S., Anand K., Tran H., Yu F., Finefield J.M., Sunderhaus J.D.,
RA   McAfoos T.J., Tsukamoto S., Williams R.M., Sherman D.H.;
RT   "Comparative analysis of the biosynthetic systems for fungal
RT   bicyclo[2.2.2]diazaoctane indole alkaloids: the (+)/(-)-notoamide,
RT   paraherquamide and malbrancheamide pathways.";
RL   Med. Chem. Commun. 3:987-996(2012).
RN   [2]
RP   BIOTECHNOLOGY.
RX   PubMed=18986806; DOI=10.1016/j.bmcl.2008.10.057;
RA   Miller K.A., Figueroa M., Valente M.W., Greshock T.J., Mata R.,
RA   Williams R.M.;
RT   "Calmodulin inhibitory activity of the malbrancheamides and various
RT   analogs.";
RL   Bioorg. Med. Chem. Lett. 18:6479-6481(2008).
RN   [3]
RP   BIOTECHNOLOGY.
RX   PubMed=19185562; DOI=10.1016/j.ab.2009.01.002;
RA   Gonzalez-Andrade M., Figueroa M., Rodriguez-Sotres R., Mata R.,
RA   Sosa-Peinado A.;
RT   "An alternative assay to discover potential calmodulin inhibitors using a
RT   human fluorophore-labeled CaM protein.";
RL   Anal. Biochem. 387:64-70(2009).
RN   [4]
RP   BIOTECHNOLOGY.
RX   PubMed=20939762; DOI=10.3109/14756366.2010.518964;
RA   Figueroa M., Gonzalez-Andrade M., Sosa-Peinado A., Madariaga-Mazon A.,
RA   Del Rio-Portilla F., Gonzalez M.C., Mata R.;
RT   "Fluorescence, circular dichroism, NMR, and docking studies of the
RT   interaction of the alkaloid malbrancheamide with calmodulin.";
RL   J. Enzym. Inhib. Med. Chem. 26:378-385(2011).
RN   [5]
RP   BIOTECHNOLOGY.
RX   PubMed=25643751; DOI=10.1111/jphp.12346;
RA   Madariaga-Mazon A., Hernandez-Abreu O., Estrada-Soto S., Mata R.;
RT   "Insights on the vasorelaxant mode of action of malbrancheamide.";
RL   J. Pharm. Pharmacol. 67:551-558(2015).
RN   [6]
RP   FUNCTION.
RX   PubMed=28777910; DOI=10.1021/jacs.7b06773;
RA   Fraley A.E., Garcia-Borras M., Tripathi A., Khare D., Mercado-Marin E.V.,
RA   Tran H., Dan Q., Webb G.P., Watts K.R., Crews P., Sarpong R.,
RA   Williams R.M., Smith J.L., Houk K.N., Sherman D.H.;
RT   "Function and structure of MalA/MalA', iterative halogenases for late-stage
RT   C-H functionalization of indole alkaloids.";
RL   J. Am. Chem. Soc. 139:12060-12068(2017).
RN   [7]
RP   FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF TYR-2132, AND PATHWAY.
RX   PubMed=31548667; DOI=10.1038/s41557-019-0326-6;
RA   Dan Q., Newmister S.A., Klas K.R., Fraley A.E., McAfoos T.J., Somoza A.D.,
RA   Sunderhaus J.D., Ye Y., Shende V.V., Yu F., Sanders J.N., Brown W.C.,
RA   Zhao L., Paton R.S., Houk K.N., Smith J.L., Sherman D.H., Williams R.M.;
RT   "Fungal indole alkaloid biogenesis through evolution of a bifunctional
RT   reductase/Diels-Alderase.";
RL   Nat. Chem. 11:972-980(2019).
CC   -!- FUNCTION: Nonribisomal peptide synthase; part of the gene cluster that
CC       mediates the biosynthesis of malbrancheamide, a dichlorinated fungal
CC       indole alkaloid that belongs to a family of natural products containing
CC       a characteristic bicyclo[2.2.2]diazaoctane core (PubMed:23213353,
CC       PubMed:31548667, PubMed:28777910). The first step of malbrancheamide
CC       biosynthesis involves coupling of L-proline and L-tryptophan by malG, a
CC       bimodular NRPS, to produce L-Pro-L-Trp aldehyde through reductive
CC       offloading (PubMed:23213353, PubMed:31548667). This compound undergoes
CC       spontaneous cyclization and dehydration to give a dienamine which is
CC       reverse prenylated at C-2 by malE (PubMed:31548667). The other
CC       prenyltransferase present in the cluster, malB, displays modest
CC       activity, suggesting that may be a redundant gene in the pathway
CC       (PubMed:31548667). Subsequently, a [4+2] Diels-Alder cyclo-addition
CC       catalyzed by the bifunctional enzyme malC forms the characteristic
CC       bicyclo[2.2.2]diazaoctane ring of premalbrancheamid (PubMed:31548667).
CC       Finally, the flavin-dependent halogenase malA catalyzes the iterative
CC       dichlorination of the indole ring of premalbrancheamide to yield C-9
CC       monochlorinated malbrancheamide B, C-8 monochlorinated
CC       isomalbrancheamide B, and dichlorinated malbrancheamide
CC       (PubMed:31548667, PubMed:28777910). MalA is also able to brominate
CC       premalbrancheamide at C-9 to yield malbrancheamide C, and, to a lesser
CC       extend, at C-8 to yield isomalbrancheamide C (PubMed:28777910).
CC       Finally, malA can brominate C-9 monochlorinated malbrancheamide B at C-
CC       8 to yield malbrancheamide D, or C-8 monochlorinated isomalbrancheamide
CC       B at C-9 to produce isomalbrancheamide D (PubMed:28777910).
CC       {ECO:0000269|PubMed:23213353, ECO:0000269|PubMed:28777910,
CC       ECO:0000269|PubMed:31548667}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=2 ATP + L-proline + L-tryptophan + NADPH = (S)-3-(indol-3-
CC         ylmethyl)-6,7,8,8a-tetrahydropyrrolo[1,2-a]pyrazin-1-one + 2 AMP + 2
CC         diphosphate + H(+) + H2O + NADP(+); Xref=Rhea:RHEA:62284,
CC         ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616,
CC         ChEBI:CHEBI:33019, ChEBI:CHEBI:57783, ChEBI:CHEBI:57912,
CC         ChEBI:CHEBI:58349, ChEBI:CHEBI:60039, ChEBI:CHEBI:145652,
CC         ChEBI:CHEBI:456215; Evidence={ECO:0000269|PubMed:31548667};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62285;
CC         Evidence={ECO:0000269|PubMed:31548667};
CC   -!- DOMAIN: NRP synthetases are composed of discrete domains (adenylation
CC       (A), thiolation (T) or peptidyl carrier protein (PCP) and condensation
CC       (C) domains) which when grouped together are referred to as a single
CC       module. Each module is responsible for the recognition (via the A
CC       domain) and incorporation of a single amino acid into the growing
CC       peptide product. Thus, an NRP synthetase is generally composed of one
CC       or more modules and can terminate in a thioesterase domain (TE) that
CC       releases the newly synthesized peptide from the enzyme. Occasionally,
CC       methyltransferase domains (responsible for amino acid methylation) are
CC       present within the NRP synthetase (By similarity). MalG has the
CC       following bimodular architecture: A1-T1-C1-A2-T2-R. MalG finishes with
CC       a reductase-like domain (R) for peptide release, which is consistent
CC       with the monooxopiperazine moiety of malbrancheamide (PubMed:31548667).
CC       {ECO:0000250|UniProtKB:A0A144KPJ6, ECO:0000269|PubMed:31548667}.
CC   -!- BIOTECHNOLOGY: Malbrancheamides have the ability to inhibit calmodulin,
CC       calmodulin-dependent phosphodiesterase (PDE1), and induce both
CC       endothelium-independent and endothelium-dependent relaxant effects,
CC       suggesting their potential as vasorelaxant agents.
CC       {ECO:0000269|PubMed:18986806, ECO:0000269|PubMed:19185562,
CC       ECO:0000269|PubMed:20939762, ECO:0000269|PubMed:25643751}.
CC   -!- SIMILARITY: Belongs to the NRP synthetase family. {ECO:0000305}.
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DR   EMBL; JQ708193; AGA37267.1; -; Genomic_DNA.
DR   AlphaFoldDB; L0E2Z1; -.
DR   SMR; L0E2Z1; -.
DR   BioCyc; MetaCyc:MON-21923; -.
DR   GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR   GO; GO:0016874; F:ligase activity; IEA:UniProtKB-KW.
DR   GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR   GO; GO:0031177; F:phosphopantetheine binding; IEA:InterPro.
DR   GO; GO:0009058; P:biosynthetic process; IEA:UniProt.
DR   Gene3D; 1.10.1200.10; -; 2.
DR   Gene3D; 3.30.300.30; -; 2.
DR   Gene3D; 3.30.559.10; -; 1.
DR   Gene3D; 3.40.50.12780; -; 2.
DR   InterPro; IPR010071; AA_adenyl_domain.
DR   InterPro; IPR036736; ACP-like_sf.
DR   InterPro; IPR045851; AMP-bd_C_sf.
DR   InterPro; IPR020845; AMP-binding_CS.
DR   InterPro; IPR000873; AMP-dep_Synth/Lig.
DR   InterPro; IPR042099; ANL_N_sf.
DR   InterPro; IPR023213; CAT-like_dom_sf.
DR   InterPro; IPR001242; Condensatn.
DR   InterPro; IPR013120; Far_NAD-bd.
DR   InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR   InterPro; IPR020806; PKS_PP-bd.
DR   InterPro; IPR009081; PP-bd_ACP.
DR   InterPro; IPR006162; Ppantetheine_attach_site.
DR   Pfam; PF00501; AMP-binding; 2.
DR   Pfam; PF00668; Condensation; 1.
DR   Pfam; PF07993; NAD_binding_4; 1.
DR   Pfam; PF00550; PP-binding; 2.
DR   SMART; SM00823; PKS_PP; 2.
DR   SUPFAM; SSF47336; SSF47336; 2.
DR   SUPFAM; SSF51735; SSF51735; 1.
DR   TIGRFAMs; TIGR01733; AA-adenyl-dom; 1.
DR   PROSITE; PS00455; AMP_BINDING; 2.
DR   PROSITE; PS50075; CARRIER; 2.
DR   PROSITE; PS00012; PHOSPHOPANTETHEINE; 2.
PE   1: Evidence at protein level;
KW   Isomerase; Ligase; Multifunctional enzyme; Oxidoreductase;
KW   Phosphopantetheine; Phosphoprotein; Repeat.
FT   CHAIN           1..2345
FT                   /note="Nonribisomal peptide synthase malG"
FT                   /id="PRO_0000448778"
FT   DOMAIN          766..839
FT                   /note="Carrier 1"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT   DOMAIN          1843..1926
FT                   /note="Carrier 2"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT   REGION          226..620
FT                   /note="Adenylation 1"
FT                   /evidence="ECO:0000255"
FT   REGION          877..1292
FT                   /note="Condensation 1"
FT                   /evidence="ECO:0000255"
FT   REGION          1317..1707
FT                   /note="Adenylation 2"
FT                   /evidence="ECO:0000255"
FT   REGION          1969..2256
FT                   /note="Reductase (R) domain"
FT                   /evidence="ECO:0000255"
FT   MOD_RES         800
FT                   /note="O-(pantetheine 4'-phosphoryl)serine"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT   MOD_RES         1885
FT                   /note="O-(pantetheine 4'-phosphoryl)serine"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT   MUTAGEN         2132
FT                   /note="Y->F: Impairs the reductive release of the malG
FT                   product."
FT                   /evidence="ECO:0000269|PubMed:31548667"
SQ   SEQUENCE   2345 AA;  258201 MW;  0988DB3F35FD67F9 CRC64;
     MSDDPLLSSP TEAICLNHSV TDLRLAAALK LSWTFLLAHY SGSSEIPLDI RLEYRYIDGS
     ETNFEPFDAT FEVDKKSLIE DSIGMIQNML TPSTRPHSLS NGINSSQSDK HVPEAQVSFT
     FSSGSRPVLE KGATTRYAAT VLELECLQGL KKEYLCRINF NRMMWNVEEA TGILRQFRHI
     AQQIVSADVC ATLSQINLMC ESDIEQLKRW NSTVPDPVLA CIHELFSEQA KKNPTATAVQ
     TSEGSFDYGR LDELSSALAC HLSSNGLTRG TPVPLLFDKS MWMVVATLAV LKAGATCVSI
     CTGLPTKAIE DILEQTAAQL VLVSESQGLR LSETRTQVVS DKTMQIWHTM SGKPELPQSD
     PTDLACIIFT SGSTGKPKGI MLDHIALVTS IRNHGPSLGI SSSSRALQFS SYAFDMSFYE
     TYTTLLSGGC ICIPSETERL NSLPQFICDH NVNWAFLTPS VLRDFHPSEF PSLRTLATGG
     EPVGADIANE WAGRLQLFNL WGPAEATICA TGPILPGVWI PGTFGKAVGC IAWITQAENP
     DELVPIGAVG EVLIEGPVLA QGYSGDVEKT KASFIPFPKW RERFELTPRG RVLFRTGDLA
     QYNPDGTIRY VGRMGTVVKV GGQRVDIDAV EYALRRIDRS SHIAVEAVEL EKETGQGPTL
     IAFLSSDMNG VSGSEKKRCC SIDPGSRSWE AWANIAIRLQ DTLAGVLPRY MIPHLFIPVS
     TIPTTPSGKA NKRQLQALVL GQSKAHLLQL CRQRSPDASY PEQHLTENET LLRLLVSDVL
     GIDRDHVAMN SRFFHLGGDS LAAVKLVALA RQQGIQLKVE AILQSCSLRE AAGTMISAGE
     KQKLQSKTSF AINKCDDKLG LLEEATVQCG ISESDIEEIY PSTPLQEGLI TVTSTFSASK
     PYVDKILFTL SATADLDRVR DAWNHVVAAN DILRSRIILS PAGKAFNVVV RSEPSWQYYK
     TVQQYLENDN AQDMTFGKEL ITFNLIASHD QSASARSIGI TIHHALYDNW TVSLLHKQAE
     DAYRGELVEP CSFSTFSHYV LQQSPDINKE FWRKQFLDLR AGTFPELPSS DYVPRANSSS
     QHLYKGQHQR RDFSMATNIQ LAWALLLSLY TNSPDVVYGL VVNGRMAPMP GVGGLVGPTI
     ATVPFRTTVE RSMSVQAALE AIQKRVLSIV PFEQTGLQNI ARMGEGPKTA CNFQNLLVIQ
     QDLEFKGEGI FCRRQNLVGA VNNFPGYGII LLCSATEHGW AFEILYSNSL IPETRARRIL
     LQLDHLLRQL EVDPYRQLAQ LELLCPSDKS KLTSWNTQLP IRVNACIPEV FGAQCLVRSE
     RTAVSAWDGS LSYRELDRFS SIVARHLQAV GVGKGTITPI LFEKSRWVVV AMLAVLKTGA
     AFVMLDTNQP LQRKQGICRA VRATTIATSA SCAHESKVLA NSIYVLDEAS ITKTDTNQFL
     PLVEVSPNDL AYVVFTSGST GEPKGVLIEH ASSCSASRAQ AAKLGISPDS RVLQLSSYTF
     DSFAVEILAS LLAGCCICIP SESESSNDIA GAVRRFSATW LCITPSVLGL TNPDEVPSLK
     TVVAVGESAR PSQIRLWSTR VNFICGYGPS ECSTGASAQL IRSAGSDPRI IGSGMGSCLW
     VAHTDDHNVL VPIGAIGELL IQGPIVGRGY MNSPEKTRAA FLESTAWIPE FRQVATERFY
     KTGDLVRQNE DGSIVYLGRK NREVKLRGQR LDLEEVENQL SAALEMDINI VAEVVKPKGV
     DSQPVLIAFF QVVADVELRS DNITFLELNP DIGLRLLDAE EKLRKILPPV MIPSVYLQVQ
     RMPLTMSGKM NRQALRNKAS TRTLSQLFSS GSVRHEDDYL TLQPHESTAL FVCQAICGIM
     RDKIDDTKTL IAGKNVNLSR TGMDSIDAMM LARTISRHFG ITLSIRAFLG SSVTVRDIAR
     LIEGVKSEDN LSQFDLYAKY ESIWEELRGV VRGLTPSDKP QLCDKTPAGM SVFLTGGTGF
     LGTHILRQIL QDPRVELVTV LTRAESPAHA LSKIVESAKI AQWWQESYRN RIDAWVGDLA
     RPRLGLSDDH WARLCGYGEH KFTSIIHNGA AVHWGYDFEK LKPVNVMSTF WLLVSLFIAG
     PLVNFTYVSA LLPECDGLTD REIALKTSDD GYSQTKYVSE LLVKNFKEQL CNNPIAIVRP
     GLLIGSAEHG VANVGDYLWR VVSSAFSVGA YISEKGDAWI YIAAVDWVAN QVIREALYES
     TTDLRIINVT DGLTVKEFWR AIQIASPRQL NALQSEDWLS LIRQQLDVTG KSHPLWPVIS
     FLESSKGCLG FSHNLPPQAH SLSSMIITAL IKNVRYLASL GLVSWTTTGS NCDHSVQQRI
     FRRVL