PPAF2_HOLDI
ID PPAF2_HOLDI Reviewed; 415 AA.
AC Q9GRW0;
DT 28-FEB-2018, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2001, sequence version 1.
DT 03-AUG-2022, entry version 100.
DE RecName: Full=Phenoloxidase-activating factor 2 {ECO:0000305|PubMed:11012672};
DE AltName: Full=45 KDa PPAF {ECO:0000303|PubMed:11012672};
DE Short=Hd-45 {ECO:0000303|PubMed:11012672};
DE AltName: Full=Prophenoloxidase-activating factor II {ECO:0000303|PubMed:11012672};
DE AltName: Full=Serine protease-like PPAF-2 {ECO:0000305|PubMed:11012672};
DE Contains:
DE RecName: Full=Phenoloxidase-activating factor 2 light chain {ECO:0000305|PubMed:11012672, ECO:0000305|PubMed:16362048};
DE Contains:
DE RecName: Full=Phenoloxidase-activating factor 2 heavy chain {ECO:0000305|PubMed:11012672, ECO:0000305|PubMed:16362048};
DE Flags: Precursor;
GN Name=PPAF2 {ECO:0000305|PubMed:11012672};
GN Synonyms=PPAF-II {ECO:0000303|PubMed:11012672};
OS Holotrichia diomphalia (Korean black chafer).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota;
OC Neoptera; Endopterygota; Coleoptera; Polyphaga; Scarabaeiformia;
OC Scarabaeidae; Melolonthinae; Holotrichia.
OX NCBI_TaxID=33394 {ECO:0000312|EMBL:CAC12665.1};
RN [1] {ECO:0000312|EMBL:CAC12665.1}
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 100-106; 216-230; 261-268
RP AND 331-343, FUNCTION, DEVELOPMENTAL STAGE, INDUCTION, AND PROTEOLYTIC
RP CLEAVAGE.
RC TISSUE=Larva {ECO:0000312|EMBL:CAC12665.1};
RX PubMed=11012672; DOI=10.1046/j.1432-1327.2000.01695.x;
RA Kwon T.H., Kim M.S., Choi H.W., Joo C.H., Cho M.Y., Lee B.L.;
RT "A masquerade-like serine proteinase homologue is necessary for
RT phenoloxidase activity in the coleopteran insect, Holotrichia diomphalia
RT larvae.";
RL Eur. J. Biochem. 267:6188-6196(2000).
RN [2] {ECO:0000305}
RP FUNCTION, SUBUNIT, AND SUBCELLULAR LOCATION.
RX PubMed=9652393; DOI=10.1046/j.1432-1327.1998.2540050.x;
RA Lee S.Y., Kwon T.H., Hyun J.H., Choi J.S., Kawabata S.I., Iwanaga S.,
RA Lee B.L.;
RT "In vitro activation of pro-phenol-oxidase by two kinds of pro-phenol-
RT oxidase-activating factors isolated from hemolymph of coleopteran,
RT Holotrichia diomphalia larvae.";
RL Eur. J. Biochem. 254:50-57(1998).
RN [3] {ECO:0000305}
RP PROTEIN SEQUENCE OF 100-109, AND TISSUE SPECIFICITY.
RX PubMed=12185078; DOI=10.1074/jbc.m205508200;
RA Kim M.S., Baek M.J., Lee M.H., Park J.W., Lee S.Y., Soderhall K., Lee B.L.;
RT "A new easter-type serine protease cleaves a masquerade-like protein during
RT prophenoloxidase activation in Holotrichia diomphalia larvae.";
RL J. Biol. Chem. 277:39999-40004(2002).
RN [4] {ECO:0007744|PDB:2B9L}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 25-415 IN COMPLEX WITH CALCIUM,
RP FUNCTION, SUBUNIT, INTERACTION WITH PPO1 AND PPO2, PROTEOLYTIC CLEAVAGE,
RP GLYCOSYLATION AT ASN-32, DISULFIDE BONDS, AND MUTAGENESIS OF THR-60; GLN-64
RP AND VAL-111.
RX PubMed=16362048; DOI=10.1038/sj.emboj.7600891;
RA Piao S., Song Y.L., Kim J.H., Park S.Y., Park J.W., Lee B.L., Oh B.H.,
RA Ha N.C.;
RT "Crystal structure of a clip-domain serine protease and functional roles of
RT the clip domains.";
RL EMBO J. 24:4404-4414(2005).
CC -!- FUNCTION: Binds and activates processed prophenoloxidases PPO1 and PPO2
CC and thus is involved in the activation of the prophenoloxidase cascade
CC probably following the recognition of pathogen-derived products.
CC {ECO:0000269|PubMed:11012672, ECO:0000269|PubMed:12185078,
CC ECO:0000269|PubMed:16362048, ECO:0000269|PubMed:9652393,
CC ECO:0000303|PubMed:12185078}.
CC -!- SUBUNIT: Heterodimer of a light chain and a heavy chain; disulfide-
CC linked (Probable). Can oligomerize into a dodecamer (PubMed:11012672,
CC PubMed:16362048. PubMed:9652393). Interacts (via CLIP domain) with
CC cleaved PPO1 and/or PPO2; the interaction results in PPO1 and/or PPO2
CC activation (PubMed:16362048). {ECO:0000269|PubMed:11012672,
CC ECO:0000269|PubMed:16362048, ECO:0000269|PubMed:9652393,
CC ECO:0000305|PubMed:16362048}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:9652393}.
CC Note=Secreted in the hemolymph. {ECO:0000269|PubMed:9652393}.
CC -!- TISSUE SPECIFICITY: Hemocytes. {ECO:0000269|PubMed:12185078}.
CC -!- DEVELOPMENTAL STAGE: Expressed in larva (at protein level).
CC {ECO:0000269|PubMed:11012672}.
CC -!- INDUCTION: Up-regulated in response to Gram-negative bacterial
CC infections. {ECO:0000269|PubMed:11012672}.
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 (By similarity). The clip domain interacts with
CC cleaved PPO1 and/or PPO2; the interaction results in PPO1 and/or PPO2
CC activation (PubMed:16362048). {ECO:0000255|PROSITE-ProRule:PRU01236,
CC ECO:0000269|PubMed:16362048}.
CC -!- PTM: The N-terminus is blocked (PubMed:11012672). Proteolytically
CC cleaved by PPAF3 (PubMed:11012672, PubMed:12185078). Cleavage produces
CC a light chain and a heavy chain, which probably remain covalently
CC associated through an interchain disulfide bond (Probable).
CC {ECO:0000269|PubMed:11012672, ECO:0000269|PubMed:12185078,
CC ECO:0000269|PubMed:16362048, ECO:0000305|PubMed:16362048}.
CC -!- MISCELLANEOUS: Stabilized by calcium. {ECO:0000269|PubMed:16362048}.
CC -!- SIMILARITY: Belongs to the peptidase S1 family. CLIP subfamily.
CC {ECO:0000255|PROSITE-ProRule:PRU01236}.
CC -!- CAUTION: Lacks the conserved Ser residue within the catalytic triad
CC which is replaced by a Gly residue, probably resulting in a loss of
CC proteolytic activity. {ECO:0000305}.
CC -!- CAUTION: It is not clear if the light chain is degraded after cleavage.
CC {ECO:0000305}.
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DR EMBL; AJ400903; CAC12665.1; -; mRNA.
DR PDB; 2B9L; X-ray; 2.00 A; A=25-415.
DR PDBsum; 2B9L; -.
DR AlphaFoldDB; Q9GRW0; -.
DR SMR; Q9GRW0; -.
DR MEROPS; S01.960; -.
DR iPTMnet; Q9GRW0; -.
DR EvolutionaryTrace; Q9GRW0; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:InterPro.
DR GO; GO:0042742; P:defense response to bacterium; IEA:UniProtKB-KW.
DR GO; GO:0045087; P:innate immune response; IEA:UniProtKB-KW.
DR GO; GO:0006508; P:proteolysis; IEA:InterPro.
DR CDD; cd00190; Tryp_SPc; 1.
DR Gene3D; 2.40.10.10; -; 2.
DR InterPro; IPR022700; CLIP.
DR InterPro; IPR009003; Peptidase_S1_PA.
DR InterPro; IPR043504; Peptidase_S1_PA_chymotrypsin.
DR InterPro; IPR001314; Peptidase_S1A.
DR InterPro; IPR041515; PPAF-2_Clip.
DR InterPro; IPR001254; Trypsin_dom.
DR Pfam; PF18322; CLIP_1; 1.
DR Pfam; PF00089; Trypsin; 1.
DR PRINTS; PR00722; CHYMOTRYPSIN.
DR SMART; SM00020; Tryp_SPc; 1.
DR SUPFAM; SSF50494; SSF50494; 1.
DR PROSITE; PS51888; CLIP; 1.
DR PROSITE; PS50240; TRYPSIN_DOM; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic; Antimicrobial; Calcium;
KW Direct protein sequencing; Disulfide bond; Glycoprotein; Immunity;
KW Innate immunity; Metal-binding; Secreted; Serine protease homolog; Signal.
FT SIGNAL 1..24
FT /evidence="ECO:0000303|PubMed:11012672"
FT CHAIN 25..99
FT /note="Phenoloxidase-activating factor 2 light chain"
FT /evidence="ECO:0000305|PubMed:11012672,
FT ECO:0000305|PubMed:16362048"
FT /id="PRO_5004326545"
FT CHAIN 100..415
FT /note="Phenoloxidase-activating factor 2 heavy chain"
FT /evidence="ECO:0000305|PubMed:11012672,
FT ECO:0000305|PubMed:16362048"
FT /id="PRO_0000443317"
FT DOMAIN 57..113
FT /note="Clip"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000269|PubMed:11012672, ECO:0000269|PubMed:16362048"
FT DOMAIN 151..404
FT /note="Peptidase S1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00274"
FT BINDING 218
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT BINDING 220
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT BINDING 223
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT BINDING 226
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT SITE 99..100
FT /note="Cleavage"
FT /evidence="ECO:0000269|PubMed:11012672"
FT CARBOHYD 32
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT CARBOHYD 89
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT DISULFID 58..112
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000269|PubMed:16362048, ECO:0007744|PDB:2B9L"
FT DISULFID 69..105
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000269|PubMed:16362048, ECO:0007744|PDB:2B9L"
FT DISULFID 75..113
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01236,
FT ECO:0000269|PubMed:16362048, ECO:0007744|PDB:2B9L"
FT DISULFID 139..272
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT DISULFID 185..201
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT DISULFID 285..359
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT DISULFID 318..339
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT DISULFID 349..380
FT /evidence="ECO:0000269|PubMed:16362048,
FT ECO:0007744|PDB:2B9L"
FT MUTAGEN 60
FT /note="T->A: No effect on expected cleavage,
FT oligomerization and activity of PPO1 and PPO2."
FT /evidence="ECO:0000269|PubMed:16362048"
FT MUTAGEN 64
FT /note="Q->A: No effect on expected cleavage,
FT oligomerization and activity of PPO1 and PPO2."
FT /evidence="ECO:0000269|PubMed:16362048"
FT MUTAGEN 111
FT /note="V->A: Loss of interaction with PPO1 and PPO2."
FT /evidence="ECO:0000269|PubMed:16362048"
FT HELIX 25..30
FT /evidence="ECO:0007829|PDB:2B9L"
FT TURN 31..34
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 52..54
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 61..63
FT /evidence="ECO:0007829|PDB:2B9L"
FT TURN 64..66
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 67..71
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 72..74
FT /evidence="ECO:0007829|PDB:2B9L"
FT TURN 77..79
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 95..97
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 102..107
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 110..114
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 150..153
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 155..157
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 166..171
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 182..191
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 194..197
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 199..202
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 203..205
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 209..211
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 212..217
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 226..228
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 231..240
FT /evidence="ECO:0007829|PDB:2B9L"
FT TURN 246..248
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 254..260
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 285..289
FT /evidence="ECO:0007829|PDB:2B9L"
FT TURN 292..296
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 306..310
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 315..323
FT /evidence="ECO:0007829|PDB:2B9L"
FT TURN 324..327
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 337..340
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 343..345
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 356..361
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 364..375
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 382..385
FT /evidence="ECO:0007829|PDB:2B9L"
FT STRAND 387..391
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 392..395
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 396..405
FT /evidence="ECO:0007829|PDB:2B9L"
FT HELIX 411..413
FT /evidence="ECO:0007829|PDB:2B9L"
SQ SEQUENCE 415 AA; 45258 MW; 062EC180504E0125 CRC64;
MKRLFVITAF FLFGAEAQNS VIDAAVVNIF GNASEYIPPG YEIVTKAPLG ALTALPRCGT
GADQGKKVCI VYHRCDGVTN TVTPEEVINT TGEGIFDIRE NANECESYLD VCCGLPEGGV
LPTPSPTPPV VPVLKPSFCG IRNERGLDFK ITGQTNEAEY GEFPWMVAVL KANVIPGSGE
EQLVCGGSLI APSVVLTGAH CVNSYQSNLD AIKIRAGEWD TLTEKERLPY QERKIRQVII
HSNFNPKTVV NDVALLLLDR PLVQADNIGT ICLPQQSQIF DSTECFASGW GKKEFGSRHR
YSNILKKIQL PTVDRDKCQA DLRNTRLGLK FVLDQTFVCA GGEQGKDTCT GDGGSPLFCP
DPRNPSRYMQ MGIVAWGIGC GDENVPGVYA NVAHFRNWID QEMQAKGLST TPYVE
