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 07-APR-2021, entry version 46. 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)s(-1) for native protein, 0.130 M(-1)s(-1) CC for native protein with 10 mM Ca(2+) and 0.116 M(-1)s(-1) for EDTA- CC 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}. 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; 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 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 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:0007744|PDB:4TOQ" FT HELIX 37..39 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 42..47 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 52..63 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 66..70 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 78..80 FT /evidence="ECO:0007744|PDB:4TOQ" FT TURN 81..85 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 86..94 FT /evidence="ECO:0007744|PDB:4TOQ" FT TURN 95..97 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 99..105 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 116..130 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 131..133 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 146..151 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 160..168 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 173..175 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 178..181 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 184..188 FT /evidence="ECO:0007744|PDB:4TOQ" FT TURN 190..192 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 193..196 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 202..207 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 224..236 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 240..249 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 250..252 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 253..255 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 260..266 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 268..271 FT /evidence="ECO:0007744|PDB:4TOQ" FT STRAND 277..283 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 285..287 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 291..295 FT /evidence="ECO:0007744|PDB:4TOQ" FT HELIX 296..298 FT /evidence="ECO:0007744|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 //