POPB_GALM3
ID POPB_GALM3 Reviewed; 730 AA.
AC H2E7Q8; A0A067SC43;
DT 28-MAR-2018, integrated into UniProtKB/Swiss-Prot.
DT 22-JAN-2014, sequence version 2.
DT 03-AUG-2022, entry version 32.
DE RecName: Full=Dual function macrocyclase-peptidase POPB {ECO:0000303|PubMed:29051530};
DE EC=3.4.21.26 {ECO:0000269|PubMed:28866879, ECO:0000269|PubMed:29051530};
DE AltName: Full=Prolyl oligopeptidase B {ECO:0000303|PubMed:22202811};
DE Short=POP B {ECO:0000303|PubMed:22202811};
DE AltName: Full=Toxin-processing prolyl oligopeptidase {ECO:0000303|PubMed:22202811};
GN Name=POPB {ECO:0000303|PubMed:22202811}; ORFNames=GALMADRAFT_78538;
OS Galerina marginata (strain CBS 339.88).
OC Eukaryota; Fungi; Dikarya; Basidiomycota; Agaricomycotina; Agaricomycetes;
OC Agaricomycetidae; Agaricales; Strophariaceae; Galerina.
OX NCBI_TaxID=685588;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], AND FUNCTION.
RC STRAIN=CBS 339.88;
RX PubMed=22202811; DOI=10.1016/j.fgb.2011.12.005;
RA Luo H., Hallen-Adams H.E., Scott-Craig J.S., Walton J.D.;
RT "Ribosomal biosynthesis of alpha-amanitin in Galerina marginata.";
RL Fungal Genet. Biol. 49:123-129(2012).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=CBS 339.88;
RX PubMed=24958869; DOI=10.1073/pnas.1400592111;
RA Riley R., Salamov A.A., Brown D.W., Nagy L.G., Floudas D., Held B.W.,
RA Levasseur A., Lombard V., Morin E., Otillar R., Lindquist E.A., Sun H.,
RA LaButti K.M., Schmutz J., Jabbour D., Luo H., Baker S.E., Pisabarro A.G.,
RA Walton J.D., Blanchette R.A., Henrissat B., Martin F., Cullen D.,
RA Hibbett D.S., Grigoriev I.V.;
RT "Extensive sampling of basidiomycete genomes demonstrates inadequacy of the
RT white-rot/brown-rot paradigm for wood decay fungi.";
RL Proc. Natl. Acad. Sci. U.S.A. 111:9923-9928(2014).
RN [3]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=28866879; DOI=10.1021/acssynbio.7b00264;
RA Sgambelluri R.M., Smith M.O., Walton J.D.;
RT "Versatility of prolyl oligopeptidase B in peptide macrocyclization.";
RL ACS Synth. Biol. 7:145-152(2018).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.44 ANGSTROMS) IN COMPLEX WITH SUBSTRATE, FUNCTION,
RP MUTAGENESIS OF SER-577; ASP-661; ARG-663; TRP-695 AND HIS-698, ACTIVE SITE,
RP CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=29051530; DOI=10.1038/s41467-017-00862-4;
RA Czekster C.M., Ludewig H., McMahon S.A., Naismith J.H.;
RT "Characterization of a dual function macrocyclase enables design and use of
RT efficient macrocyclization substrates.";
RL Nat. Commun. 8:1045-1045(2017).
CC -!- FUNCTION: Dual function macrocyclase-peptidase involved in the
CC biosynthesis of the highly toxic amanitin toxin family of macrocycles
CC (PubMed:22202811, PubMed:28866879, PubMed:29051530). Cleaves peptide
CC bonds on the C-terminal side of prolyl residues (PubMed:29051530). The
CC enzyme first removes 10 residues from the N-terminus of a 35-residue
CC substrate (PubMed:29051530). Conformational trapping of the 25 amino-
CC acid peptide forces the enzyme to release this intermediate rather than
CC proceed to macrocyclization (PubMed:29051530). The enzyme rebinds the
CC 25 amino-acid peptide in a different conformation and catalyzes
CC macrocyclization of the N-terminal eight residues (PubMed:28866879,
CC PubMed:29051530). {ECO:0000269|PubMed:22202811,
CC ECO:0000269|PubMed:28866879, ECO:0000269|PubMed:29051530}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of Pro-|-Xaa >> Ala-|-Xaa in oligopeptides.;
CC EC=3.4.21.26; Evidence={ECO:0000269|PubMed:28866879,
CC ECO:0000269|PubMed:29051530};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=23 uM for the full alpha-amanitin 35mer proprotein
CC {ECO:0000269|PubMed:29051530};
CC KM=51 uM for the 25mer IWGIGCNPWTAEHVDQTLASGNDIC
CC {ECO:0000269|PubMed:29051530};
CC KM=19 uM for the 25mer LWGIGCNPWTAEHVDQTLASGNDIC
CC {ECO:0000269|PubMed:29051530};
CC KM=8 uM for the 25mer IFGIGCNPWTAEHVDQTLASGNDIC
CC {ECO:0000269|PubMed:29051530};
CC KM=28 uM for the 25mer IWGIGSNPWTAEHVDQTLASGNDIC
CC {ECO:0000269|PubMed:29051530};
CC KM=10 uM for the 25mer IWGIGCNPWTAEHVDQTLASGNDIS
CC {ECO:0000269|PubMed:29051530};
CC KM=25 uM for the 19mer IWGIGCNPDQTLASGNDIC
CC {ECO:0000269|PubMed:29051530};
CC KM=380 uM for the 14mer IWGIGCNPWTAEHV {ECO:0000269|PubMed:29051530};
CC KM=24.4 uM for the 13mer IWGIGCNPWTAEH {ECO:0000269|PubMed:29051530};
CC -!- SUBUNIT: Monomer. {ECO:0000250|UniProtKB:P48147}.
CC -!- SIMILARITY: Belongs to the peptidase S9A family. {ECO:0000305}.
CC -!- WEB RESOURCE: Name=Protein Spotlight; Note=More to it - Issue 202 of
CC April 2018;
CC URL="https://web.expasy.org/spotlight/back_issues/202/";
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DR EMBL; JN827314; AEX26938.2; -; mRNA.
DR EMBL; KL142408; KDR68475.1; -; Genomic_DNA.
DR PDB; 5N4B; X-ray; 1.44 A; A/B=1-730.
DR PDB; 5N4C; X-ray; 2.19 A; A/B/C/D=1-730.
DR PDB; 5N4D; X-ray; 1.62 A; A/B=1-730.
DR PDB; 5N4E; X-ray; 2.90 A; A/B=1-730.
DR PDB; 5N4F; X-ray; 2.40 A; A=1-730.
DR PDBsum; 5N4B; -.
DR PDBsum; 5N4C; -.
DR PDBsum; 5N4D; -.
DR PDBsum; 5N4E; -.
DR PDBsum; 5N4F; -.
DR AlphaFoldDB; H2E7Q8; -.
DR SMR; H2E7Q8; -.
DR ESTHER; 9agar-h2e7q8; S9N_PPCE_Peptidase_S9.
DR MEROPS; S09.077; -.
DR EnsemblFungi; KDR68475; KDR68475; GALMADRAFT_78538.
DR OrthoDB; 225446at2759; -.
DR Proteomes; UP000027222; Unassembled WGS sequence.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProt.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:UniProtKB-EC.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR002471; Pept_S9_AS.
DR InterPro; IPR023302; Pept_S9A_N.
DR InterPro; IPR001375; Peptidase_S9.
DR InterPro; IPR002470; Peptidase_S9A.
DR Pfam; PF00326; Peptidase_S9; 1.
DR Pfam; PF02897; Peptidase_S9_N; 1.
DR PRINTS; PR00862; PROLIGOPTASE.
DR SUPFAM; SSF53474; SSF53474; 1.
DR PROSITE; PS00708; PRO_ENDOPEP_SER; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Protease; Reference proteome; Serine protease.
FT CHAIN 1..730
FT /note="Dual function macrocyclase-peptidase POPB"
FT /id="PRO_0000443717"
FT REGION 1..34
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 1..20
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 577
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10084,
FT ECO:0000269|PubMed:29051530"
FT ACT_SITE 661
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10084,
FT ECO:0000269|PubMed:29051530"
FT ACT_SITE 698
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10084,
FT ECO:0000269|PubMed:29051530"
FT MUTAGEN 577
FT /note="S->A: Impairs catalytic activity but still binds
FT bith 35mer and 25mer substrates."
FT /evidence="ECO:0000269|PubMed:29051530"
FT MUTAGEN 661
FT /note="D->A: Impairs catalytic activity but still binds
FT bith 35mer and 25mer substrates."
FT /evidence="ECO:0000269|PubMed:29051530"
FT MUTAGEN 663
FT /note="R->A,K,Q: Leads to diminished activities for both
FT peptide bond hydrolysis and macrocyclization."
FT /evidence="ECO:0000269|PubMed:29051530"
FT MUTAGEN 695
FT /note="Missing: Leads to diminished activities for both
FT peptide bond hydrolysis and macrocyclization."
FT /evidence="ECO:0000269|PubMed:29051530"
FT MUTAGEN 698
FT /note="H->A: Impairs catalytic activity but still binds
FT bith 35mer and 25mer substrates."
FT /evidence="ECO:0000269|PubMed:29051530"
FT STRAND 20..25
FT /evidence="ECO:0007829|PDB:5N4B"
FT TURN 26..28
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 29..34
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 38..41
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 45..63
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 67..79
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 95..100
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 102..106
FT /evidence="ECO:0007829|PDB:5N4F"
FT STRAND 108..116
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 120..122
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 124..127
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 128..132
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 134..136
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 143..150
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 154..163
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 167..175
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 182..187
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 197..200
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 211..218
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 235..240
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 245..247
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 249..253
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 261..266
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 272..283
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 286..291
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 304..307
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 309..312
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 314..320
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 323..328
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 336..341
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 344..346
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 348..353
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 359..367
FT /evidence="ECO:0007829|PDB:5N4B"
FT TURN 368..370
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 371..378
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 381..387
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 393..398
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 403..408
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 413..422
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 425..433
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 438..440
FT /evidence="ECO:0007829|PDB:5N4F"
FT STRAND 442..447
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 455..457
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 458..466
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 472..480
FT /evidence="ECO:0007829|PDB:5N4B"
FT TURN 485..488
FT /evidence="ECO:0007829|PDB:5N4F"
FT STRAND 491..494
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 509..518
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 521..525
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 535..539
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 543..546
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 547..562
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 571..576
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 578..589
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 596..602
FT /evidence="ECO:0007829|PDB:5N4B"
FT TURN 607..609
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 610..612
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 616..619
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 620..623
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 629..635
FT /evidence="ECO:0007829|PDB:5N4B"
FT TURN 636..638
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 640..642
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 646..648
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 652..658
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 662..664
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 667..679
FT /evidence="ECO:0007829|PDB:5N4B"
FT STRAND 687..692
FT /evidence="ECO:0007829|PDB:5N4B"
FT HELIX 704..721
FT /evidence="ECO:0007829|PDB:5N4B"
SQ SEQUENCE 730 AA; 81781 MW; 354827749ED68A4B CRC64;
MSSVTWAPGN YPSTRRSDHV DTYQSASKGE VPVPDPYQWL EESTDEVDKW TTAQADLAQS
YLDQNADIQK LAEKFRASRN YAKFSAPTLL DDGHWYWFYN RGLQSQSVLY RSKEPALPDF
SKGDDNVGDV FFDPNVLAAD GSAGMVLCKF SPDGKFFAYA VSHLGGDYST IYVRSTSSPL
SQASVAQGVD GRLSDEVKWF KFSTIIWTKD SKGFLYQRYP ARERHEGTRS DRNAMMCYHK
VGTTQEEDII VYQDNEHPEW IYGADTSEDG KYLYLYQFKD TSKKNLLWVA ELDEDGVKSG
IHWRKVVNEY AADYNIITNH GSLVYIKTNL NAPQYKVITI DLSKDEPEIR DFIPEEKDAK
LAQVNCANEE YFVAIYKRNV KDEIYLYSKA GVQLTRLAPD FVGAASIANR QKQTHFFLTL
SGFNTPGTIA RYDFTAPETQ RFSILRTTKV NELDPDDFES TQVWYESKDG TKIPMFIVRH
KSTKFDGTAA AIQYGYGGFA TSADPFFSPI ILTFLQTYGA IFAVPSIRGG GEFGEEWHKG
GRRETKVNTF DDFIAAAQFL VKNKYAAPGK VAINGASNGG LLVMGSIVRA PEGTFGAAVP
EGGVADLLKF HKFTGGQAWI SEYGNPSIPE EFDYIYPLSP VHNVRTDKVM PATLITVNIG
DGRVVPMHSF KFIATLQHNV PQNPHPLLIK IDKSWLGHGM GKPTDKNVKD AADKWGFIAR
ALGLELKTVE
