PPAE_BOMMO
ID PPAE_BOMMO Reviewed; 441 AA.
AC Q9XXV0; H9JW33;
DT 28-FEB-2018, integrated into UniProtKB/Swiss-Prot.
DT 28-FEB-2018, sequence version 2.
DT 03-AUG-2022, entry version 116.
DE RecName: Full=Phenoloxidase-activating enzyme {ECO:0000305};
DE EC=3.4.21.- {ECO:0000269|PubMed:10066809, ECO:0000269|PubMed:4197814, ECO:0000269|PubMed:4197815, ECO:0000269|PubMed:7981662};
DE AltName: Full=Prophenoloxidase-activating enzyme {ECO:0000303|PubMed:4197814};
DE Contains:
DE RecName: Full=Phenoloxidase-activating enzyme light chain {ECO:0000303|PubMed:10066809};
DE Contains:
DE RecName: Full=Phenoloxidase-activating enzyme heavy chain {ECO:0000303|PubMed:10066809};
DE Flags: Precursor;
GN Name=PPAE {ECO:0000303|PubMed:4197814};
OS Bombyx mori (Silk moth).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota;
OC Neoptera; Endopterygota; Lepidoptera; Glossata; Ditrysia; Bombycoidea;
OC Bombycidae; Bombycinae; Bombyx.
OX NCBI_TaxID=7091 {ECO:0000312|EMBL:BAA76308.1};
RN [1] {ECO:0000312|EMBL:BAA76308.1}
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 22-32 AND 174-189,
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT,
RP DEVELOPMENTAL STAGE, GLYCOSYLATION, PROTEOLYTIC CLEAVAGE, MASS
RP SPECTROMETRY, DISULFIDE BONDS, AND PYROGLUTAMATE FORMATION AT GLN-22.
RC TISSUE=Integument {ECO:0000312|EMBL:BAA76308.1};
RX PubMed=10066809; DOI=10.1074/jbc.274.11.7441;
RA Satoh D., Horii A., Ochiai M., Ashida M.;
RT "Prophenoloxidase-activating enzyme of the silkworm, Bombyx mori.
RT Purification, characterization, and cDNA cloning.";
RL J. Biol. Chem. 274:7441-7453(1999).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=p50T;
RX PubMed=19121390; DOI=10.1016/j.ibmb.2008.11.004;
RG International Silkworm Genome Consortium;
RT "The genome of a lepidopteran model insect, the silkworm Bombyx mori.";
RL Insect Biochem. Mol. Biol. 38:1036-1045(2008).
RN [3] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND DEVELOPMENTAL STAGE.
RX PubMed=4197814; DOI=10.1016/0003-9861(73)90406-2;
RA Dohke K.;
RT "Studies on prephenoloxidase-activating enzyme from cuticle of the silkworm
RT Bombyx mori. I. Activation reaction by the enzyme.";
RL Arch. Biochem. Biophys. 157:203-209(1973).
RN [4] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND DEVELOPMENTAL STAGE.
RX PubMed=4197815; DOI=10.1016/0003-9861(73)90407-4;
RA Dohke K.;
RT "Studies on prephenoloxidase-activating enzyme from cuticle of the silkworm
RT Bombyx mori. II. Purification and characterization of the enzyme.";
RL Arch. Biochem. Biophys. 157:210-221(1973).
RN [5] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, AND DEVELOPMENTAL STAGE.
RX PubMed=7981662;
RA Aso Y., Yamashita T., Meno K., Murakami M.;
RT "Inhibition of prophenoloxidase-activating enzyme from Bombyx mori by
RT endogenous chymotrypsin inhibitors.";
RL Biochem. Mol. Biol. Int. 33:751-758(1994).
CC -!- FUNCTION: Endopeptidase with selective post-Arg cleavage site
CC (PubMed:4197814, PubMed:4197815, PubMed:10066809, PubMed:7981662).
CC Activates prophenoloxidase (PubMed:4197814, PubMed:4197815). Has a
CC probable role in the melanization process as part of the innate immune
CC response (Probable). {ECO:0000269|PubMed:10066809,
CC ECO:0000269|PubMed:4197814, ECO:0000269|PubMed:4197815,
CC ECO:0000269|PubMed:7981662, ECO:0000305}.
CC -!- ACTIVITY REGULATION: Stabilized by calcium (PubMed:4197814). Inhibited
CC by di-isopropyl phosphorofluoridate (DFP),
CC phenylmethanesulfonylfluoride (PMSF), p-nitrophenyl-p'-
CC guanidinobenzonate (p-NPGB), p-chloromercuribenzoate (PCMB),
CC ethylenediaminetetraacetic acid (EDTA), urea and CI-13c
CC (PubMed:4197814, PubMed:4197815, PubMed:7981662).
CC {ECO:0000269|PubMed:4197814, ECO:0000269|PubMed:4197815,
CC ECO:0000269|PubMed:7981662}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.2 uM for prophenoloxidase (at pH 7.6 and 0 degrees Celsius)
CC {ECO:0000269|PubMed:4197814, ECO:0000269|PubMed:4197815};
CC KM=1.43 mM for N-benzoyl-L-arginine ethyl ester hydrochloride (BAEE)
CC (in the absence of KCl) {ECO:0000269|PubMed:4197815};
CC KM=0.77 mM for N-benzoyl-L-arginine ethyl ester hydrochloride (BAEE)
CC (in the presence of 0.025 M KCl) {ECO:0000269|PubMed:4197815};
CC KM=1.2 mM for N-benzoyl-L-arginine ethyl ester hydrochloride (BAEE)
CC (in the presence of 0.004 M KCl) {ECO:0000269|PubMed:10066809};
CC KM=0.35 mM for N-benzoyl-L-arginine ethyl ester hydrochloride (BAEE)
CC (in the presence of 0.5 M KCl) {ECO:0000269|PubMed:10066809};
CC KM=0.145 mM for N-benzoyl-L-arginine ethyl ester hydrochloride (BAEE)
CC (in the presence of 0.5 M KCl) {ECO:0000269|PubMed:4197815};
CC KM=0.709 mM for N-p-tosyl-L-arginine methyl ester hydrochloride
CC (TAME) (in the presence of 0.5 M KCl) {ECO:0000269|PubMed:4197815};
CC pH dependence:
CC Optimum pH is 6.0-8.5. {ECO:0000269|PubMed:10066809};
CC Temperature dependence:
CC Optimum temperature is 0-25 degrees Celsius.
CC {ECO:0000269|PubMed:10066809};
CC -!- SUBUNIT: In the active form, heterodimer of a light chain and a heavy
CC chain; disulfide-linked. {ECO:0000305|PubMed:10066809}.
CC -!- DEVELOPMENTAL STAGE: Expressed in eggs and larval cuticles (at protein
CC level) (PubMed:10066809, PubMed:4197814, PubMed:4197815,
CC PubMed:7981662). Expressed in larval hemocytes and salivary glands
CC (PubMed:10066809). {ECO:0000269|PubMed:10066809,
CC ECO:0000269|PubMed:4197814, ECO:0000269|PubMed:4197815,
CC ECO:0000269|PubMed:7981662}.
CC -!- DOMAIN: The clip domain consists of 35-55 residues which are 'knitted'
CC together usually by 3 conserved disulfide bonds forming a clip-like
CC compact structure. {ECO:0000255|PROSITE-ProRule:PRU01236}.
CC -!- PTM: Proteolytically cleaved for activation (PubMed:10066809). Cleavage
CC produces a light chain and a catalytic heavy chain which remains
CC covalently associated probably through an interchain disulfide bond
CC (Probable). {ECO:0000269|PubMed:10066809, ECO:0000305|PubMed:10066809}.
CC -!- PTM: Glycosylated. {ECO:0000269|PubMed:10066809}.
CC -!- MASS SPECTROMETRY: [Phenoloxidase-activating enzyme heavy chain]:
CC Mass=32848; Method=MALDI; Evidence={ECO:0000269|PubMed:10066809};
CC -!- SIMILARITY: Belongs to the peptidase S1 family. CLIP subfamily.
CC {ECO:0000255|PROSITE-ProRule:PRU01236}.
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; AB009670; BAA76308.1; -; mRNA.
DR EMBL; BABH01025554; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR RefSeq; NP_001036832.1; NM_001043367.1.
DR AlphaFoldDB; Q9XXV0; -.
DR SMR; Q9XXV0; -.
DR STRING; 7091.BGIBMGA013746-TA; -.
DR MEROPS; S01.305; -.
DR GeneID; 692368; -.
DR KEGG; bmor:692368; -.
DR CTD; 692368; -.
DR eggNOG; KOG3627; Eukaryota.
DR HOGENOM; CLU_006842_0_3_1; -.
DR OrthoDB; 1314811at2759; -.
DR Proteomes; UP000005204; Unassembled WGS sequence.
DR GO; GO:0004175; F:endopeptidase activity; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008233; F:peptidase activity; IDA:UniProtKB.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:InterPro.
DR GO; GO:0016485; P:protein processing; IDA:UniProtKB.
DR CDD; cd00190; Tryp_SPc; 1.
DR Gene3D; 2.40.10.10; -; 2.
DR Gene3D; 3.30.1640.30; -; 2.
DR InterPro; IPR022700; CLIP.
DR InterPro; IPR038565; CLIP_sf.
DR InterPro; IPR009003; Peptidase_S1_PA.
DR InterPro; IPR043504; Peptidase_S1_PA_chymotrypsin.
DR InterPro; IPR001314; Peptidase_S1A.
DR InterPro; IPR001254; Trypsin_dom.
DR InterPro; IPR018114; TRYPSIN_HIS.
DR InterPro; IPR033116; TRYPSIN_SER.
DR Pfam; PF12032; CLIP; 2.
DR Pfam; PF00089; Trypsin; 1.
DR PRINTS; PR00722; CHYMOTRYPSIN.
DR SMART; SM00680; CLIP; 2.
DR SMART; SM00020; Tryp_SPc; 1.
DR SUPFAM; SSF50494; SSF50494; 1.
DR PROSITE; PS51888; CLIP; 2.
DR PROSITE; PS50240; TRYPSIN_DOM; 1.
DR PROSITE; PS00134; TRYPSIN_HIS; 1.
DR PROSITE; PS00135; TRYPSIN_SER; 1.
PE 1: Evidence at protein level;
KW Calcium; Direct protein sequencing; Disulfide bond; Glycoprotein;
KW Hydrolase; Metal-binding; Protease; Pyrrolidone carboxylic acid;
KW Reference proteome; Serine protease; Signal; Zymogen.
FT SIGNAL 1..21
FT /evidence="ECO:0000269|PubMed:10066809"
FT CHAIN 22..173
FT /note="Phenoloxidase-activating enzyme light chain"
FT /evidence="ECO:0000305|PubMed:10066809"
FT /id="PRO_5004338749"
FT CHAIN 174..441
FT /note="Phenoloxidase-activating enzyme heavy chain"
FT /evidence="ECO:0000305|PubMed:10066809"
FT /id="PRO_0000443318"
FT DOMAIN 23..76
FT /note="Clip 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000303|PubMed:10066809"
FT DOMAIN 77..127
FT /note="Clip 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000303|PubMed:10066809"
FT DOMAIN 174..440
FT /note="Peptidase S1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00274"
FT ACT_SITE 218
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00274"
FT ACT_SITE 285
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00274"
FT ACT_SITE 391
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00274"
FT BINDING 237
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250|UniProtKB:O97366"
FT BINDING 239
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250|UniProtKB:O97366"
FT BINDING 242
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250|UniProtKB:O97366"
FT BINDING 246
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250|UniProtKB:O97366"
FT SITE 173..174
FT /note="Cleavage"
FT /evidence="ECO:0000269|PubMed:10066809"
FT MOD_RES 22
FT /note="Pyrrolidone carboxylic acid"
FT /evidence="ECO:0000269|PubMed:10066809"
FT CARBOHYD 239
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 334
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT DISULFID 24..75
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000303|PubMed:10066809"
FT DISULFID 34..65
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000303|PubMed:10066809"
FT DISULFID 40..76
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000303|PubMed:10066809"
FT DISULFID 78..126
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000303|PubMed:10066809"
FT DISULFID 88..117
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000303|PubMed:10066809"
FT DISULFID 94..127
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000303|PubMed:10066809"
FT DISULFID 164..305
FT /note="Interchain (between light and heavy chains)"
FT /evidence="ECO:0000250|UniProtKB:Q9VB68"
FT DISULFID 203..219
FT /evidence="ECO:0000250|UniProtKB:Q9VB68"
FT DISULFID 356..377
FT /evidence="ECO:0000250|UniProtKB:Q9VB68"
FT DISULFID 387..416
FT /evidence="ECO:0000250|UniProtKB:Q9VB68"
FT CONFLICT 223
FT /note="A -> G (in Ref. 1; BAA76308)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 441 AA; 47985 MW; D503F4BBE64FFBA2 CRC64;
MFLIWTFIVA VLAIQTKSVV AQSCRTPNGL NGNCVSVYEC QALLAILNNQ RRTQQDEKFL
RDSQCGTKNS VPAVCCPCNA ADGQQGNCVN INSCPYVLQL LKNPNEANLN YVRGSVCQGS
EQQSICCVTA PQSTAVTTTP RPKRVHACQS EMTATPPNPE GKCCGRDIAV GDKIVGGAPA
SIDSYPWLVV IEYVRLERTM LLCGGALISG KYVLTAGHCV KGAILDVGTP KTVRLGEYNT
TNPGRDCVSV SAGGTDCTDP LVKIGIEKTI PHPDYQPYHF LRKHDIGLIR LQSIAPFTDF
IRPICLPSTD YTVNPPSKFA LTVAGWGRYL QFDNGTVRSS KIKLHVTLPF VQRDVCEANQ
KPLRNGQRIT LWKGQMCAGG EAGKDSCKGD SGGPLMYEHS KKYEAVGIVS FGPEKCGQID
IPGVYTNVYE YLPWIQNTIE P
