MALC_MALAU
ID MALC_MALAU Reviewed; 264 AA.
AC L0E4F8;
DT 11-DEC-2019, integrated into UniProtKB/Swiss-Prot.
DT 06-MAR-2013, sequence version 1.
DT 03-AUG-2022, entry version 27.
DE RecName: Full=Short-chain dehydrogenase/reductase malC {ECO:0000303|PubMed:23213353};
DE EC=1.1.-.- {ECO:0000269|PubMed:31548667};
DE AltName: Full=Malbrancheamide biosynthesis cluster protein C {ECO:0000303|PubMed:23213353};
GN Name=malC {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], AND FUNCTION.
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 X-RAY CRYSTALLOGRAPHY (1.6 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY,
RP COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, 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: Short-chain dehydrogenase/reductase; part of the gene cluster
CC that mediates the biosynthesis of malbrancheamide, a dichlorinated
CC fungal indole alkaloid that belongs to a family of natural products
CC containing a characteristic bicyclo[2.2.2]diazaoctane core
CC (PubMed:23213353, PubMed:28777910, PubMed:31548667). The first step of
CC malbrancheamide biosynthesis involves coupling of L-proline and L-
CC tryptophan by malG, a bimodular NRPS, to produce L-Pro-L-Trp aldehyde
CC through reductive offloading (PubMed:23213353, PubMed:31548667). This
CC compound undergoes spontaneous cyclization and dehydration to give a
CC dienamine which is reverse prenylated at C-2 by malE (PubMed:31548667).
CC The other prenyltransferase present in the cluster, malB, displays
CC modest 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 The first reaction catalyzed is a NADPH-dependent reduction reaction in
CC which the nicotinamide cofactor is a stoichiometric reagent
CC (PubMed:31548667). Either NADH or NADPH is effective as a cofactor
CC (PubMed:31548667). Finally, the flavin-dependent halogenase malA
CC catalyzes the iterative dichlorination of the indole ring of
CC premalbrancheamide to yield C-9 monochlorinated malbrancheamide B, C-8
CC monochlorinated isomalbrancheamide B, and dichlorinated malbrancheamide
CC (PubMed:28777910, PubMed:31548667). 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=1-hydroxy-3-{[2-(1,1-dimethylallyl)-indol-3-yl]methyl}-
CC 6H,7H,8H-5lambda(5)-pyrrolo[1,2-a]pyrazine + H(+) + NADPH = 1-
CC hydroxy-3-{[2-(1,1-dimethylallyl)-indol-3-yl]methyl}-4H,6H,7H,8H-
CC pyrrolo[1,2-a]pyrazine + NADP(+); Xref=Rhea:RHEA:62300,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:145657, ChEBI:CHEBI:145675;
CC Evidence={ECO:0000269|PubMed:31548667};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62301;
CC Evidence={ECO:0000269|PubMed:31548667};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=1-hydroxy-3-{[2-(1,1-dimethylallyl)-indol-3-yl]methyl}-
CC 4H,6H,7H,8H-pyrrolo[1,2-a]pyrazine = (+)-premalbrancheamide;
CC Xref=Rhea:RHEA:62304, ChEBI:CHEBI:145658, ChEBI:CHEBI:145675;
CC Evidence={ECO:0000269|PubMed:31548667};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62305;
CC Evidence={ECO:0000269|PubMed:31548667};
CC -!- COFACTOR:
CC Name=NADP(+); Xref=ChEBI:CHEBI:58349; Evidence={ECO:0000305};
CC Note=NADP(+) is required for stereocontrolled formation of
CC premalbrancheamide, however it does not appear to be required as a
CC formal stoichiometric reagent because the second reaction performed by
CC malC, the [4+2] cycloaddition, is a balanced chemical reaction without
CC requirement for hydride transfer to balance the reaction (Probable).
CC {ECO:0000269|PubMed:31548667, ECO:0000305};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=16 uM for NADPH {ECO:0000269|PubMed:31548667};
CC KM=93 uM for NADH {ECO:0000269|PubMed:31548667};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000269|PubMed:31548667}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
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 short-chain dehydrogenases/reductases (SDR)
CC family. {ECO:0000305}.
CC -!- CAUTION: MalC lacks the characteristic Tyr and Lys catalytic amino
CC acids, and also the essential Asn and Ser residues of typical SDR
CC family proteins, suggesting that the active site is reconfigured to fit
CC its unique catalytic roles. {ECO:0000269|PubMed:31548667}.
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DR EMBL; JQ708193; AGA37263.1; -; Genomic_DNA.
DR PDB; 6NKH; X-ray; 1.60 A; A/B/C/D=1-264.
DR PDBsum; 6NKH; -.
DR AlphaFoldDB; L0E4F8; -.
DR SMR; L0E4F8; -.
DR BioCyc; MetaCyc:MON-21925; -.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR002347; SDR_fam.
DR PRINTS; PR00081; GDHRDH.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Glycoprotein; Membrane; NADP; Oxidoreductase; Transmembrane;
KW Transmembrane helix.
FT CHAIN 1..264
FT /note="Short-chain dehydrogenase/reductase malC"
FT /id="PRO_0000448776"
FT TRANSMEM 13..35
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 17..41
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:P50162"
FT BINDING 22
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 23
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 25
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 45
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 46
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 49
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 75
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 130
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 202
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT BINDING 204
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:L0E2Z4"
FT CARBOHYD 249
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT HELIX 5..11
FT /evidence="ECO:0007829|PDB:6NKH"
FT STRAND 15..19
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 24..35
FT /evidence="ECO:0007829|PDB:6NKH"
FT STRAND 39..45
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 47..58
FT /evidence="ECO:0007829|PDB:6NKH"
FT STRAND 69..73
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 79..94
FT /evidence="ECO:0007829|PDB:6NKH"
FT STRAND 101..104
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 115..117
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 120..130
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 132..140
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 141..143
FT /evidence="ECO:0007829|PDB:6NKH"
FT STRAND 151..156
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 159..161
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 169..188
FT /evidence="ECO:0007829|PDB:6NKH"
FT TURN 189..191
FT /evidence="ECO:0007829|PDB:6NKH"
FT STRAND 192..199
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 205..211
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 212..214
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 215..225
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 234..246
FT /evidence="ECO:0007829|PDB:6NKH"
FT STRAND 254..258
FT /evidence="ECO:0007829|PDB:6NKH"
FT HELIX 261..263
FT /evidence="ECO:0007829|PDB:6NKH"
SQ SEQUENCE 264 AA; 28233 MW; C8A236ADC06B0B1B CRC64;
MAPTRRSRDL LRGKNVLIIG GTSGIGFAVA QLVIEHGAMA CIAGSNPTKL GKALDALKQH
PDRDPIAIVQ SATCDLFDVP NLEQNLDNLL KLAAGDSKIH HIVFTAADMV QPPPLASVTI
EQIQRVGTIR FTAPMLVAKL LPKYMELCPE NSYTLTSGSH AKQPDPGWSL VTGYCGGVEG
LMRGLAVDMM PLRVNVVSPG AVLTPVLRDI LGDSLEIALD AARKKSTTGR IARPEDVAEA
YLYIMKDQNI TGTVLETSAG MLLR
