PHQB_PENFE
ID PHQB_PENFE Reviewed; 2449 AA.
AC L0E2U2;
DT 26-FEB-2020, 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 phqB {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:0000305|PubMed:23213353};
DE AltName: Full=Paraherquamide biosynthesis cluster protein B {ECO:0000303|PubMed:23213353};
GN Name=phqB {ECO:0000303|PubMed:23213353};
OS Penicillium fellutanum.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium.
OX NCBI_TaxID=70095;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, DOMAIN, AND PATHWAY.
RC STRAIN=ATCC 20841 / MF5123;
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] {ECO:0007744|PDB:6NKI}
RP X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS) OF 2016-2449 IN COMPLEX WITH NADPH,
RP FUNCTION, CATALYTIC ACTIVITY, 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 paraherquamide, a fungal indole alkaloid
CC that belongs to a family of natural products containing a
CC characteristic bicyclo[2.2.2]diazaoctane core (PubMed:23213353). The
CC first steps in the biosynthesis of paraherquamide is the production of
CC the beta-methyl-proline precursor from L-isoleucine (Probable). They
CC require oxidation of a terminally hydroxylated L-isoleucine to the
CC corresponding aldehyde by enzymes which have still to be identified
CC (Probable). Spontaneous cyclization and dehydration would yield the 4-
CC methyl pyrolline-5-carboxylic acid, which is then reduced by the
CC pyrroline-5-carboxylate reductase phqD leading to the beta-methyl-
CC proline precursor (Probable). The next step of paraherquamide
CC biosynthesis involves coupling of beta-methyl-proline and L-tryptophan
CC by the bimodular NRPS phqB, to produce a monooxopiperazine intermediate
CC (Probable). The reductase (R) domain of phqB utilizes NADPH for hydride
CC transfer to reduce the thioester bond of the T domain-tethered linear
CC dipeptide to a hemithioaminal intermediate, which spontaneously cleaves
CC the C-S bond to release the aldehyde product (PubMed:31548667). This
CC compound undergoes spontaneous cyclization and dehydration to give a
CC dienamine which is reverse prenylated at C-2 by the reverse
CC prenyltransferase phqJ (Probable). The other prenyltransferase present
CC in the cluster, phqI may be a redundant gene in the pathway (Probable).
CC During biosynthetic assembly, the key step to produce the polycyclic
CC core is catalyzed by the bifunctional reductase and intramolecular
CC [4+2] Diels-Alderase, phqE, resulting in formation of the [2.2.2]
CC diazaoctane intermediate preparaherquamide (PubMed:31548667). Following
CC formation of preparaherquamide, an indole 2,3-epoxidation-initiated
CC pinacol-like rearrangement is catalyzed by the phqK FAD-dependent
CC monooxygenase (Probable). The prenyltransferase phqA, the cytochrome
CC P450 monooxygenase phqL, and the FAD-linked oxidoreductase phqH (or the
CC cytochrome P450 monooxygenase phqM), are proposed to be involved in the
CC formation of the pyran ring (Probable). The FAD-dependent monooxygenase
CC phqK is likely responsible for generation of the spiro-oxindole, and
CC the N-methylation is likely mediated by the phqN methyltransferase
CC leading to the isolable natural product paraherquamide F (Probable).
CC However, the order of these biosynthetic steps has still to be
CC determined (Probable). In late-stage paraherquamide biosynthesis, the
CC third P450 monooxygenase, phqO, is probably responsible for the C-14
CC hydroxylation, transforming paraherquamide F to paraherquamide G, and
CC paraherquamide E to the final product paraherquamide A (Probable). The
CC expansion from the 6-membered ring pyran (in paraherquamides F and G)
CC to the 7-membered dioxepin ring (in paraherquamides A and E) represents
CC a poorly understood but intriguing process that probably involves the
CC 2-oxoglutarate-dependent dioxygenase phqC (Probable). Finally, the
CC remaining members of the paraherquamide cluster, including phqI as well
CC as phqM (or phqH), do not have a clearly prescribed role and appear to
CC be redundant (Probable). {ECO:0000269|PubMed:23213353,
CC ECO:0000269|PubMed:31548667, ECO:0000305|PubMed:23213353}.
CC -!- PATHWAY: Alkaloid biosynthesis. {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). PhqB has the
CC following architecture: A1-T1-C1-A2-T2-R. PhqB finishes with a
CC reductase-like domain (R) for peptide release, which is consistent with
CC the monooxopiperazine moiety of paraherquamide (Probable).
CC {ECO:0000250|UniProtKB:A0A144KPJ6, ECO:0000305|PubMed:23213353}.
CC -!- SIMILARITY: Belongs to the NRP synthetase family. {ECO:0000305}.
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DR EMBL; JQ708195; AGA37269.1; -; Genomic_DNA.
DR PDB; 6NKI; X-ray; 2.60 A; A=2016-2449.
DR PDBsum; 6NKI; -.
DR AlphaFoldDB; L0E2U2; -.
DR SMR; L0E2U2; -.
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:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR Gene3D; 1.10.1200.10; -; 1.
DR Gene3D; 3.30.300.30; -; 2.
DR Gene3D; 3.30.559.10; -; 1.
DR Gene3D; 3.40.50.12780; -; 2.
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; IPR006162; Ppantetheine_attach_site.
DR Pfam; PF00501; AMP-binding; 2.
DR Pfam; PF00668; Condensation; 1.
DR Pfam; PF07993; NAD_binding_4; 1.
DR SUPFAM; SSF47336; SSF47336; 2.
DR SUPFAM; SSF51735; SSF51735; 1.
DR PROSITE; PS00455; AMP_BINDING; 1.
DR PROSITE; PS00012; PHOSPHOPANTETHEINE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; Isomerase; Ligase;
KW Multifunctional enzyme; Oxidoreductase; Phosphopantetheine; Phosphoprotein;
KW Repeat.
FT CHAIN 1..2449
FT /note="Nonribisomal peptide synthase phqB"
FT /id="PRO_0000448866"
FT DOMAIN 795..870
FT /note="Carrier 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT DOMAIN 1915..1993
FT /note="Carrier 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT REGION 253..654
FT /note="Adenylation 1"
FT /evidence="ECO:0000255"
FT REGION 913..1337
FT /note="Condensation 1"
FT /evidence="ECO:0000255"
FT REGION 1357..1756
FT /note="Adenylation 2"
FT /evidence="ECO:0000255"
FT REGION 2041..2297
FT /note="Reductase (R) domain"
FT /evidence="ECO:0000255, ECO:0000269|PubMed:31548667"
FT BINDING 2045
FT /ligand="NADPH"
FT /ligand_id="ChEBI:CHEBI:57783"
FT /evidence="ECO:0007744|PDB:6NKI"
FT BINDING 2249
FT /ligand="NADPH"
FT /ligand_id="ChEBI:CHEBI:57783"
FT /evidence="ECO:0007744|PDB:6NKI"
FT BINDING 2259
FT /ligand="NADPH"
FT /ligand_id="ChEBI:CHEBI:57783"
FT /evidence="ECO:0007744|PDB:6NKI"
FT MOD_RES 815
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT MOD_RES 1952
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT HELIX 2019..2028
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2037..2042
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2047..2058
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2059..2073
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2074..2087
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2093..2098
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2099..2103
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2109..2112
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2115..2118
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2137..2140
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2151..2171
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2177..2181
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2217..2235
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2241..2245
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2248..2250
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2263..2274
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2276..2278
FT /evidence="ECO:0007829|PDB:6NKI"
FT TURN 2280..2283
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2284..2290
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2291..2303
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2325..2327
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2331..2333
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2334..2345
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2350..2352
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2354..2366
FT /evidence="ECO:0007829|PDB:6NKI"
FT TURN 2367..2371
FT /evidence="ECO:0007829|PDB:6NKI"
FT TURN 2373..2377
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2378..2384
FT /evidence="ECO:0007829|PDB:6NKI"
FT STRAND 2394..2396
FT /evidence="ECO:0007829|PDB:6NKI"
FT HELIX 2404..2425
FT /evidence="ECO:0007829|PDB:6NKI"
SQ SEQUENCE 2449 AA; 268893 MW; 72EC9B1B0943597D CRC64;
MAKQGCIRVS VPFPLVYAPT TSDSTNLRAS HQGRTTRLPL GSSLSEEYCL ISWALLASLY
HGSATVLLNG VLPSELGSPC EPCADAIVDM APDSEIQSLE RQMADFLQQI KHIPAESDAA
NDNPNVVLDF GERTASLLMQ SVPSSHSATE ESFPPIHDGE GLHLRIICQP SEPDISWFCE
VVHDENLVDR DAAQRIAEQF AHIYKQLGSH SRGLQASIND LSLLNTADEH QLIEWNHDPP
AIIDACFQDL FAIQVHSGPG RLAVSAWDGE LTYGQLDELA NQSAQRLLEQ NVRPGMVVPL
LFEKSKWMAV AMLAVAKVRA TAVCICISHP MDLMKRILYQ SNPAIILLSR AQEPLIRQIG
EYPVLIIPED LYSSPRSATA EQPVSTAFPS ASSDDVAFIV FSSGSTGVPK GIVLSHRAIA
TAGHYVGDRL QVTSEARVLQ FSSYAFDMSI IETWQALTRG ACLCIPSETQ RLNSMPEFIQ
QHRVTWAFFT PTTLRNFEPS DFPTLETLTL GGETIPVDLA RHWESRVSIF NLWGPAEVGP
AGAGPISPTS GWIPGTFGTA AGCISWITMP DDSDSLAPIG TVGEMIIEGA VVADGYLNDP
TRTQQVFIEA PRWRERFTEI PVQCRFFRTG DLCHYNPDGT LRYVGRRDTV VKIRGQRVDV
DAVELQLRQL DPHVDSVVSA VKFYDAAGAM TLVQFLVDRT GFEIDWSSQR PGESMTALES
GSWALSHHYQ SLLRPYLPQY MVPSLIIPVK SLPRTATDKV DRRRLCQCFQ QFSSSQLMDW
LGGNRSRCLA KADSLPMTPN EDVLVKVIAS VCKISTSSID LKASFPQLGG DSTTAIRLTR
VLLAYNLLLH TERLLDIECS LRTVAKEARP TDAVTLADGP PPFSLIGVND ANAITCLRRV
ASQECKIQEE DIQDIYPTTP LQEGLLAVTE IHSGDAYVDR VLFSLPADCS VELAQQAWQN
VVQATSILRT RIIQADDGRT YQIVVRPERK IQWQTASSES QFYEKDRARS MGLGSPLVRL
TLIQDSEARE QPAKLAVTFH HSVYDAFTLH ACIKQAEKAY TSETLFPSTF TGFINHLNQQ
KLADGERTRQ FWLKEMVELQ SNVFPALPSP QYLPHTSTSV VYEGCRANAT SSKQISSPSA
KVRLAWALLI SLYTDSPDIV YGTVVDGRRG LGVILGSVLG PTIATLPVRT TIQRESTVEE
SLAQVQENMK RMIPFEHTGL QRIRNMGHGP ATACKFQNLL VIQADDMIPD SPIFGPVEVS
VGSINSFPGY ALILQVAPSE TSWKFEMLVD EAVVPREQAE LMLSQLSHLL KQIDDCHTQN
LTIAQLDLIS DRDSELLSTC LKSIPTCLDS TIVDLVEAQV TRNPSKCAVS ACDGDLSYAE
LQSSARQLAQ LLLPLIVGQG IQFIPIFLER SYWVPISMLA VAKLGVAFVL LDPNQPHERN
VKICRAISGT LGITSAQMQN LASTVCDGPW ISLSTEALIS HAQAVPSGTT TTMPPMPNPS
PRDLLYAAFT SGSTGEPKAV LIEHASYASA VIAQQNKLEI TSSSRVLQLS SYAFDSFAVE
ILTVLASGGC VCIPSESEIA EDLGHVVEKY RSNWLCITPS VLRLLTPDDV PSLRTVVAVG
ESMLPGQIKL WCSRVHLYCG YGPTECCTGA AVHRVTSTDA DARLIGKGMG AVLWVVDKED
VTRRMPVNTV GELILQGPIV GRGYLNNPKK STECFLQPPS WAPQFKDRQA SRMYRTGDMV
RRNLDGTFTF LGRTNQHTKL HGQRIDLAEI ERHVLRFFGT DASGIAAILQ PTKSDMPPCL
VAMVHIPSLA AKVPDISDMN GFSNIAFQSH SHDFALRASR VQQKLRQSFP PVMVPELYLQ
LPSIPLTISG KVNRRSLLDE ATELSPVDLH DLGGLSRNES RSTGLLDHTD EPVAWALSKH
IGQLLQRKTG HEKMAAEIVG RNVGLSRVGL DSIDIIALSQ FISRHYDCSI SMTNLFDSTL
TVRMVAEMID RTPNSVPEKA LLSPGWWERV QCMIRQINDL PVCQSSRRTI HSRPSGKRLF
LTGATGFLGT HILHQLLVDN DVSIVYVLAR APCPRKGLAR IIQAARLARW WRNDYRRLIQ
VWPGDLSQPH LGLADEHWET LSGTESSLNS SIGAVDAIIH CGAVIHWGYD YDTLEAANVR
STFDILQCLN RSPTPIALTY ISALIPGDAA LATTDTDNHP SMSNGHAVFP PIELTDGYTQ
TKFASEQLIG AFSARHKAHS LTIVRPGFMI GPVSNAVANG DDLLWRVVTT AMTTCSYNSD
ESDNWLFVAA VDWVASLIIH ETLHARPSSH SVNDNANPLA PSAKAVSIGD GLNMSDFWKA
IMLGLGRDLI PSSSQRWMDT VEQQVNEVGT SHPLWPLMGF LRASGGCLGV APTDPLPVPI
YQPPSLTNMI RQAVVRNAEY LASLEDLAAS TMLFKRRNKV ALGNGLINS
