ID   NEP2_NEPGR              Reviewed;         438 AA.
AC   Q766C2;
DT   15-MAR-2005, integrated into UniProtKB/Swiss-Prot.
DT   05-JUL-2004, sequence version 1.
DT   03-AUG-2022, entry version 69.
DE   RecName: Full=Aspartic proteinase nepenthesin-2;
DE            EC=3.4.23.12;
DE   AltName: Full=Nepenthesin-II;
DE   Flags: Precursor;
GN   Name=nep2;
OS   Nepenthes gracilis (Slender pitcher plant).
OC   Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC   Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC   Caryophyllales; Nepenthaceae; Nepenthes.
OX   NCBI_TaxID=150966;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA], 3D-STRUCTURE MODELING, AND CHARACTERIZATION.
RC   TISSUE=Pitcher;
RX   PubMed=15035659; DOI=10.1042/bj20031575;
RA   Athauda S.B.P., Matsumoto K., Rajapakshe S., Kuribayashi M., Kojima M.,
RA   Kubomura-Yoshida N., Iwamatsu A., Shibata C., Inoue H., Takahashi K.;
RT   "Enzymic and structural characterization of nepenthesin, a unique member of
RT   a novel subfamily of aspartic proteinases.";
RL   Biochem. J. 381:295-306(2004).
CC   -!- FUNCTION: Extracellular proteinase found in the pitcher fluid of
CC       carnivorous plants. Digest prey for nitrogen uptake.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Similar to pepsin, but also cleaves on either side of Asp and
CC         at Lys-|-Arg.; EC=3.4.23.12;
CC   -!- ACTIVITY REGULATION: Inhibited by pepstatin and by diazoacetyl-D,L-
CC       norleucine methyl ester (DAN) in the presence of Cu(2+) ions.
CC       {ECO:0000250}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       pH dependence:
CC         Optimum pH is 3.0. Retains 80% and 60% of the original activity after
CC         incubation for 30 days at pH 3.0 and pH 4.0 respectively. Unstable at
CC         pH higher than 5.0.;
CC       Temperature dependence:
CC         Optimum temperature is 45 degrees Celsius. Thermostable up to 50
CC         degrees Celsius. Retains 44% of the original activity after
CC         incubation for 30 days at 50 degrees Celsius.;
CC   -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250}.
CC   -!- SIMILARITY: Belongs to the peptidase A1 family. {ECO:0000305}.
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DR   EMBL; AB114915; BAD07475.1; -; mRNA.
DR   AlphaFoldDB; Q766C2; -.
DR   SMR; Q766C2; -.
DR   MEROPS; A01.040; -.
DR   BRENDA; 3.4.23.12; 8734.
DR   GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR   GO; GO:0004190; F:aspartic-type endopeptidase activity; IEA:UniProtKB-KW.
DR   GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR   CDD; cd05476; pepsin_A_like_plant; 1.
DR   Gene3D; 2.40.70.10; -; 2.
DR   InterPro; IPR001461; Aspartic_peptidase_A1.
DR   InterPro; IPR001969; Aspartic_peptidase_AS.
DR   InterPro; IPR034161; Pepsin-like_plant.
DR   InterPro; IPR033121; PEPTIDASE_A1.
DR   InterPro; IPR021109; Peptidase_aspartic_dom_sf.
DR   InterPro; IPR032799; TAXi_C.
DR   InterPro; IPR032861; TAXi_N.
DR   Pfam; PF14541; TAXi_C; 1.
DR   Pfam; PF14543; TAXi_N; 1.
DR   PRINTS; PR00792; PEPSIN.
DR   SUPFAM; SSF50630; SSF50630; 1.
DR   PROSITE; PS00141; ASP_PROTEASE; 1.
DR   PROSITE; PS51767; PEPTIDASE_A1; 1.
PE   1: Evidence at protein level;
KW   Aspartyl protease; Disulfide bond; Glycoprotein; Hydrolase; Protease;
KW   Secreted; Signal; Zymogen.
FT   SIGNAL          1..24
FT                   /evidence="ECO:0000255"
FT   PROPEP          25..79
FT                   /note="Activation peptide"
FT                   /evidence="ECO:0000250"
FT                   /id="PRO_0000025916"
FT   CHAIN           80..438
FT                   /note="Aspartic proteinase nepenthesin-2"
FT                   /id="PRO_0000025917"
FT   DOMAIN          96..431
FT                   /note="Peptidase A1"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01103"
FT   ACT_SITE        114
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU10094"
FT   ACT_SITE        315
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU10094"
FT   CARBOHYD        54
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000255"
FT   DISULFID        124..127
FT                   /evidence="ECO:0000305"
FT   DISULFID        130..204
FT                   /evidence="ECO:0000305"
FT   DISULFID        151..169
FT                   /evidence="ECO:0000305"
FT   DISULFID        156..164
FT                   /evidence="ECO:0000305"
FT   DISULFID        241..435
FT                   /evidence="ECO:0000305"
FT   DISULFID        354..395
FT                   /evidence="ECO:0000305"
SQ   SEQUENCE   438 AA;  46174 MW;  91C2A79C28123A50 CRC64;
     MASPLYSVVL GLAIVSAIVA PTSSTSRGTL LHHGQKRPQP GLRVDLEQVD SGKNLTKYEL
     IKRAIKRGER RMRSINAMLQ SSSGIETPVY AGDGEYLMNV AIGTPDSSFS AIMDTGSDLI
     WTQCEPCTQC FSQPTPIFNP QDSSSFSTLP CESQYCQDLP SETCNNNECQ YTYGYGDGST
     TQGYMATETF TFETSSVPNI AFGCGEDNQG FGQGNGAGLI GMGWGPLSLP SQLGVGQFSY
     CMTSYGSSSP STLALGSAAS GVPEGSPSTT LIHSSLNPTY YYITLQGITV GGDNLGIPSS
     TFQLQDDGTG GMIIDSGTTL TYLPQDAYNA VAQAFTDQIN LPTVDESSSG LSTCFQQPSD
     GSTVQVPEIS MQFDGGVLNL GEQNILISPA EGVICLAMGS SSQLGISIFG NIQQQETQVL
     YDLQNLAVSF VPTQCGAS