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 03-JUL-2019, entry version 40. 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; Magnoliophyta; eudicotyledons; Gunneridae; OC Pentapetalae; 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 RP AND 286-296, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, SUBCELLULAR LOCATION, TISSUE RP SPECIFICITY, DEVELOPMENTAL STAGE, MASS SPECTROMETRY, AND CIRCULAR RP DICHROISM 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 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 RT analysis of sensitization patterns to chitinases."; RL Mol. Immunol. 103:89-95(2018). RN [4] RP 3D-STRUCTURE MODELING, AND SUBCELLULAR LOCATION. RX PubMed=22112454; RA Lv C., Masuda T., Yang H., Sun L., Zhao G.; RT "High-capacity calcium-binding chitinase III from pomegranate seeds RT (Punica granatum Linn.) is located in amyloplasts."; RL Plant Signal. Behav. 6:1963-1965(2011). RN [5] {ECO:0000244|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 RT the insight into its metal storage capacity."; RL Biosci. Biotechnol. Biochem. 79:45-50(2015). CC -!- FUNCTION: Hydrolyzes chitin. Probable calcium storage protein of CC the seeds. Binds calcium ions with high capacity and low affinity. CC Involved 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(- CC 1)s(-1) for native protein with 10 mM Ca(2+) and 0.116 M(-1)s(- CC 1) for EDTA-treated protein, using 4-methylumbelliferyl-beta-D- CC N,N',N''-triacetylchitotrioside (4-MU(GlcNAc)(3)) as substrate CC at pH 4.5 and 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 CC protein 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 CC of Ca(2+) from the protein by EDTA treatment decreases the CC optimum temperature to 35 degrees Celsius. CC {ECO:0000269|PubMed:21790816}; CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:21790816}. CC -!- SUBCELLULAR LOCATION: Plastid, amyloplast CC {ECO:0000269|PubMed:21790816, ECO:0000269|PubMed:22112454}. CC Note=Localizes to stroma in amyloplasts of the embryonic cells of CC the seed. {ECO:0000269|PubMed:21790816, CC ECO:0000269|PubMed:22112454}. CC -!- TISSUE SPECIFICITY: Highly expressed in seeds and to a lesser CC extent in the skin of the pomegranate fruit (at protein level). CC Not expressed in 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 CC imbibition. {ECO:0000269|PubMed:21790816}. CC -!- MASS SPECTROMETRY: Mass=29008.70; Method=MALDI; Range=27-299; CC Note=The measured mass is that of EDTA-treated protein to remove CC Ca(2+).; Evidence={ECO:0000269|PubMed:21790816}; CC -!- MASS SPECTROMETRY: Mass=29389.80; Method=MALDI; Range=27-299; CC Note=The measured mass is that of protein without EDTA-treatment CC to retain Ca(2+).; Evidence={ECO:0000269|PubMed:21790816}; CC -!- MASS SPECTROMETRY: Mass=29001.76; Method=MALDI; Range=27-299; CC Note=The measured mass is that of protein with three disulfide CC bridges.; Evidence={ECO:0000269|PubMed:30241023}; CC -!- ALLERGEN: Causes an allergic reaction in human. Binds to IgE in CC 19% of 357 patients allergic to pomegranate. CC {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 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; CHITINASE_18; 1. PE 1: Evidence at protein level; KW 3D-structure; Allergen; Amyloplast; Calcium; Carbohydrate metabolism; KW Chitin degradation; Direct protein sequencing; Disulfide bond; KW Glycosidase; Hydrolase; Metal-binding; Plastid; KW Polysaccharide degradation; Transit peptide. FT TRANSIT 1 26 Amyloplast. {ECO:0000269|PubMed:21790816, FT ECO:0000269|PubMed:30241023}. FT CHAIN 27 299 Acidic endochitinase Pun g 14, FT amyloplastic. FT {ECO:0000305|PubMed:21790816, FT ECO:0000305|PubMed:30241023}. FT /FTId=PRO_5011206581. FT ACT_SITE 153 153 Proton donor. {ECO:0000255|PROSITE- FT ProRule:PRU10053, FT ECO:0000305|PubMed:25252615}. FT DISULFID 46 93 {ECO:0000244|PDB:4TOQ, FT ECO:0000269|PubMed:25252615}. FT DISULFID 76 83 {ECO:0000244|PDB:4TOQ, FT ECO:0000269|PubMed:25252615}. FT DISULFID 185 216 {ECO:0000244|PDB:4TOQ, FT ECO:0000269|PubMed:25252615}. FT CONFLICT 90 90 I -> L (in Ref. 1; AA sequence). FT {ECO:0000305}. FT CONFLICT 295 295 I -> L (in Ref. 1; AA sequence). FT {ECO:0000305}. FT STRAND 28 35 {ECO:0000244|PDB:4TOQ}. FT HELIX 37 39 {ECO:0000244|PDB:4TOQ}. FT HELIX 42 47 {ECO:0000244|PDB:4TOQ}. FT STRAND 52 63 {ECO:0000244|PDB:4TOQ}. FT STRAND 66 70 {ECO:0000244|PDB:4TOQ}. FT HELIX 78 80 {ECO:0000244|PDB:4TOQ}. FT TURN 81 85 {ECO:0000244|PDB:4TOQ}. FT HELIX 86 94 {ECO:0000244|PDB:4TOQ}. FT TURN 95 97 {ECO:0000244|PDB:4TOQ}. FT STRAND 99 105 {ECO:0000244|PDB:4TOQ}. FT HELIX 116 130 {ECO:0000244|PDB:4TOQ}. FT STRAND 131 133 {ECO:0000244|PDB:4TOQ}. FT STRAND 146 151 {ECO:0000244|PDB:4TOQ}. FT HELIX 160 168 {ECO:0000244|PDB:4TOQ}. FT STRAND 173 175 {ECO:0000244|PDB:4TOQ}. FT STRAND 178 181 {ECO:0000244|PDB:4TOQ}. FT STRAND 184 188 {ECO:0000244|PDB:4TOQ}. FT TURN 190 192 {ECO:0000244|PDB:4TOQ}. FT HELIX 193 196 {ECO:0000244|PDB:4TOQ}. FT STRAND 202 207 {ECO:0000244|PDB:4TOQ}. FT HELIX 224 236 {ECO:0000244|PDB:4TOQ}. FT STRAND 240 249 {ECO:0000244|PDB:4TOQ}. FT HELIX 250 252 {ECO:0000244|PDB:4TOQ}. FT STRAND 253 255 {ECO:0000244|PDB:4TOQ}. FT HELIX 260 266 {ECO:0000244|PDB:4TOQ}. FT HELIX 268 271 {ECO:0000244|PDB:4TOQ}. FT STRAND 277 283 {ECO:0000244|PDB:4TOQ}. FT HELIX 285 287 {ECO:0000244|PDB:4TOQ}. FT HELIX 291 295 {ECO:0000244|PDB:4TOQ}. FT HELIX 296 298 {ECO:0000244|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 //