CHIT_PUNGR
ID CHIT_PUNGR Reviewed; 299 AA.
AC G1UH28;
DT 08-MAY-2019, integrated into UniProtKB/Swiss-Prot.
DT 16-NOV-2011, sequence version 1.
DT 25-MAY-2022, entry version 49.
DE RecName: Full=Acidic endochitinase Pun g 14, amyloplastic {ECO:0000305};
DE EC=3.2.1.14 {ECO:0000255|PROSITE-ProRule:PRU10053, ECO:0000269|PubMed:21790816, ECO:0000312|EMBL:BAK68869.1};
DE AltName: Full=Chitinase III {ECO:0000303|PubMed:21790816, ECO:0000303|PubMed:22112454, ECO:0000303|PubMed:30241023};
DE AltName: Full=Pomegranate seed chitinase {ECO:0000303|PubMed:21790816};
DE AltName: Allergen=Pun g 14 {ECO:0000303|PubMed:30241023};
DE Flags: Precursor;
GN Name=PSC {ECO:0000303|PubMed:21790816, ECO:0000312|EMBL:BAK68869.1};
GN ORFNames=CDL15_Pgr011527 {ECO:0000312|EMBL:OWM64072.1};
OS Punica granatum (Pomegranate).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; malvids; Myrtales; Lythraceae; Punica.
OX NCBI_TaxID=22663 {ECO:0000312|EMBL:BAK68869.1};
RN [1] {ECO:0000312|EMBL:BAK68869.1}
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 27-41; 67-91; 277-285 AND
RP 286-296, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, SUBCELLULAR LOCATION, TISSUE
RP SPECIFICITY, DEVELOPMENTAL STAGE, MASS SPECTROMETRY, AND CIRCULAR DICHROISM
RP ANALYSIS.
RC TISSUE=Seed {ECO:0000303|PubMed:21790816};
RX PubMed=21790816; DOI=10.1111/j.1365-313x.2011.04727.x;
RA Yang H., Zhang T., Masuda T., Lv C., Sun L., Qu G., Zhao G.;
RT "Chitinase III in pomegranate seeds (Punica granatum Linn.): a high-
RT capacity calcium-binding protein in amyloplasts.";
RL Plant J. 68:765-776(2011).
RN [2] {ECO:0000312|EMBL:OWM64072.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. Dabenzi; TISSUE=Leaf {ECO:0000312|EMBL:OWM64072.1};
RX PubMed=28654223; DOI=10.1111/tpj.13625;
RA Qin G., Xu C., Ming R., Tang H., Guyot R., Kramer E.M., Hu Y., Yi X.,
RA Qi Y., Xu X., Gao Z., Pan H., Jian J., Tian Y., Yue Z., Xu Y.;
RT "The pomegranate (Punica granatum L.) genome and the genomics of
RT punicalagin biosynthesis.";
RL Plant J. 91:1108-1128(2017).
RN [3]
RP PROTEIN SEQUENCE OF 27-37, MASS SPECTROMETRY, AND ALLERGEN.
RX PubMed=30241023; DOI=10.1016/j.molimm.2018.09.009;
RA Tuppo L., Giangrieco I., Alessandri C., Ricciardi T., Rafaiani C.,
RA Ciancamerla M., Ferrara R., Zennaro D., Bernardi M.L., Tamburrini M.,
RA Mari A., Ciardiello M.A.;
RT "Pomegranate chitinase III: Identification of a new allergen and analysis
RT of sensitization patterns to chitinases.";
RL Mol. Immunol. 103:89-95(2018).
RN [4]
RP 3D-STRUCTURE MODELING, AND SUBCELLULAR LOCATION.
RX PubMed=22112454; DOI=10.4161/psb.6.12.18147;
RA Lv C., Masuda T., Yang H., Sun L., Zhao G.;
RT "High-capacity calcium-binding chitinase III from pomegranate seeds (Punica
RT granatum Linn.) is located in amyloplasts.";
RL Plant Signal. Behav. 6:1963-1965(2011).
RN [5] {ECO:0007744|PDB:4TOQ}
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF 27-299, AND DISULFIDE BONDS.
RX PubMed=25252615; DOI=10.1080/09168451.2014.962475;
RA Masuda T., Zhao G., Mikami B.;
RT "Crystal structure of class III chitinase from pomegranate provides the
RT insight into its metal storage capacity.";
RL Biosci. Biotechnol. Biochem. 79:45-50(2015).
CC -!- FUNCTION: Hydrolyzes chitin. Probable calcium storage protein of the
CC seeds. Binds calcium ions with high capacity and low affinity. Involved
CC in seed germination. {ECO:0000269|PubMed:21790816}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Random endo-hydrolysis of N-acetyl-beta-D-glucosaminide
CC (1->4)-beta-linkages in chitin and chitodextrins.; EC=3.2.1.14;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU10053,
CC ECO:0000269|PubMed:21790816};
CC -!- ACTIVITY REGULATION: Activity is not affected by addition of 10 mM
CC Ca(2+) or removal of Ca(2+). {ECO:0000269|PubMed:21790816}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC Note=kcat/KM is 0.119 M(-1)sec(-1) for native protein, 0.130 M(-
CC 1)sec(-1) for native protein with 10 mM Ca(2+) and 0.116 M(-1)sec(-1)
CC for EDTA-treated protein, using 4-methylumbelliferyl-beta-D-N,N',N''-
CC triacetylchitotrioside (4-MU(GlcNAc)(3)) as substrate at pH 4.5 and
CC 37 degrees Celsius. {ECO:0000269|PubMed:21790816};
CC pH dependence:
CC Optimum pH is between 2.5 and 5.0. Removal of Ca(2+) from the protein
CC by EDTA treatment narrows the pH range to 2.5-3.5.
CC {ECO:0000269|PubMed:21790816};
CC Temperature dependence:
CC Optimum temperature is approximately 45 degrees Celsius. Removal of
CC Ca(2+) from the protein by EDTA treatment decreases the optimum
CC temperature to 35 degrees Celsius. {ECO:0000269|PubMed:21790816};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:21790816}.
CC -!- SUBCELLULAR LOCATION: Plastid, amyloplast {ECO:0000269|PubMed:21790816,
CC ECO:0000269|PubMed:22112454}. Note=Localizes to stroma in amyloplasts
CC of the embryonic cells of the seed. {ECO:0000269|PubMed:21790816,
CC ECO:0000269|PubMed:22112454}.
CC -!- TISSUE SPECIFICITY: Highly expressed in seeds and to a lesser extent in
CC the skin of the pomegranate fruit (at protein level). Not expressed in
CC leaves or flesh of the fruit (at protein level).
CC {ECO:0000269|PubMed:21790816}.
CC -!- DEVELOPMENTAL STAGE: Expressed at all stages of seed germination.
CC Expression decreases markedly from day 1 to day 15 after imbibition.
CC {ECO:0000269|PubMed:21790816}.
CC -!- MASS SPECTROMETRY: Mass=29008.70; Method=MALDI; Note=The measured mass
CC is that of EDTA-treated protein to remove Ca(2+).;
CC Evidence={ECO:0000269|PubMed:21790816};
CC -!- MASS SPECTROMETRY: Mass=29389.80; Method=MALDI; Note=The measured mass
CC is that of protein without EDTA-treatment to retain Ca(2+).;
CC Evidence={ECO:0000269|PubMed:21790816};
CC -!- MASS SPECTROMETRY: Mass=29001.76; Method=MALDI; Note=The measured mass
CC is that of protein with three disulfide bridges.;
CC Evidence={ECO:0000269|PubMed:30241023};
CC -!- ALLERGEN: Causes an allergic reaction in human. Binds to IgE in 19% of
CC 357 patients allergic to pomegranate. {ECO:0000269|PubMed:30241023}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 18 family. Chitinase
CC class III subfamily. {ECO:0000305}.
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DR EMBL; AB605773; BAK68869.1; -; mRNA.
DR EMBL; MTKT01005817; OWM64072.1; -; Genomic_DNA.
DR PDB; 4TOQ; X-ray; 1.60 A; A/B/C/D=27-299.
DR PDBsum; 4TOQ; -.
DR AlphaFoldDB; G1UH28; -.
DR SMR; G1UH28; -.
DR Allergome; 11786; Pun g 14.
DR Allergome; 12339; Pun g 14.0101.
DR Proteomes; UP000197138; Unassembled WGS sequence.
DR GO; GO:0009501; C:amyloplast; IDA:UniProtKB.
DR GO; GO:0005509; F:calcium ion binding; IDA:UniProtKB.
DR GO; GO:0004568; F:chitinase activity; IDA:UniProtKB.
DR GO; GO:0006032; P:chitin catabolic process; IDA:UniProtKB.
DR GO; GO:0000272; P:polysaccharide catabolic process; IEA:UniProtKB-KW.
DR GO; GO:0009845; P:seed germination; IEP:UniProtKB.
DR GO; GO:0090351; P:seedling development; IEP:UniProtKB.
DR CDD; cd02877; GH18_hevamine_XipI_class_III; 1.
DR InterPro; IPR045321; Cts1-like.
DR InterPro; IPR001223; Glyco_hydro18_cat.
DR InterPro; IPR001579; Glyco_hydro_18_chit_AS.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR Pfam; PF00704; Glyco_hydro_18; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
DR PROSITE; PS01095; GH18_1; 1.
DR PROSITE; PS51910; GH18_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Allergen; Amyloplast; Calcium; Carbohydrate metabolism;
KW Chitin degradation; Direct protein sequencing; Disulfide bond; Glycosidase;
KW Hydrolase; Metal-binding; Plastid; Polysaccharide degradation;
KW Transit peptide.
FT TRANSIT 1..26
FT /note="Amyloplast"
FT /evidence="ECO:0000269|PubMed:21790816,
FT ECO:0000269|PubMed:30241023"
FT CHAIN 27..299
FT /note="Acidic endochitinase Pun g 14, amyloplastic"
FT /evidence="ECO:0000305|PubMed:21790816,
FT ECO:0000305|PubMed:30241023"
FT /id="PRO_5011206581"
FT DOMAIN 27..299
FT /note="GH18"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01258"
FT ACT_SITE 153
FT /note="Proton donor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01258,
FT ECO:0000305|PubMed:25252615"
FT DISULFID 46..93
FT /evidence="ECO:0000269|PubMed:25252615,
FT ECO:0007744|PDB:4TOQ"
FT DISULFID 76..83
FT /evidence="ECO:0000269|PubMed:25252615,
FT ECO:0007744|PDB:4TOQ"
FT DISULFID 185..216
FT /evidence="ECO:0000269|PubMed:25252615,
FT ECO:0007744|PDB:4TOQ"
FT CONFLICT 90
FT /note="I -> L (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 295
FT /note="I -> L (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT STRAND 28..35
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 37..39
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 42..47
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 52..63
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 66..70
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 78..80
FT /evidence="ECO:0007829|PDB:4TOQ"
FT TURN 81..85
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 86..94
FT /evidence="ECO:0007829|PDB:4TOQ"
FT TURN 95..97
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 99..105
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 116..130
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 131..133
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 146..151
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 160..168
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 173..175
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 178..181
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 184..188
FT /evidence="ECO:0007829|PDB:4TOQ"
FT TURN 190..192
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 193..196
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 202..207
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 224..236
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 240..249
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 250..252
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 253..255
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 260..266
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 268..271
FT /evidence="ECO:0007829|PDB:4TOQ"
FT STRAND 277..283
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 285..287
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 291..295
FT /evidence="ECO:0007829|PDB:4TOQ"
FT HELIX 296..298
FT /evidence="ECO:0007829|PDB:4TOQ"
SQ SEQUENCE 299 AA; 31747 MW; D5D28157AA92D0B1 CRC64;
MAKTLPFSRA LLLSLSILLV ARAISAGDIA IYWGQNGGEG TLASTCDTGR YAYVIVSFVT
TFGNFRAPVV NLAGHCDPAA GTCTGLSDEI RSCQGKDIKV LMSIGGGAGD YSLVSEADAD
NFADYLWNNF LGGQSSSRPL GDAVLDGIDF DIELGTTTFY DTLARALSSR STQAAKVYLT
AAPQCPHPDS HLDAALNTGL FDNVWIQFYN NPLAQCQYSS GNTNDILSSW NTWTSSTTAG
KIFLGLPAAP EAAGSGYIPP DVLTGQILPQ IKTSAKYGGV MLYSKFYDTT YSTTIKDQV
