| IED ID | IndEnz0001000337 |
| Enzyme Type ID | amylase000337 |
| Protein Name |
Pullulanase A EC 3.2.1.41 Alpha-dextrin endo-1,6-alpha-glucosidase Pullulan 6-glucanohydrolase |
| Gene Name | spuA SP_0268 |
| Organism | Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus Streptococcus pneumoniae Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4) |
| Enzyme Sequence | MRKTPSHTEKKMVYSIRSLKNGTGSVLIGASLVLLAMATPTISSDESTPTTNEPNNRNTTTLAQPLTDTAAGSGKNESDISSPGNANASLEKTEEKPAASPADPAPQTGQDRSSEPTTSTSPVTTETKAEEPIEDNYFRIHVKKLPEENKDAQGLWTWDDVEKPSENWPNGALSFKDAKKDDYGYYLDVKLKGEQAKKISFLINNTAGKNLTGDKSVEKLVPKMNEAWLDQDYKVFSYEPQPAGTVRVNYYRTDGNYDKKSLWYWGDVKNPSSAQWPDGTDFTATGKYGRYIDIPLNEAAREFGFLLLDESKQGDDVKIRKENYKFTDLKNHSQIFLKDDDESIYTNPYYVHDIRMTGAQHVGTSSIESSFSTLVGAKKEDILKHSNITNHLGNKVTITDVAIDEAGKKVTYSGDFSDTKHPYTVSYNSDQFTTKTSWRLKDETYSYDGKLGADLKEEGKQVDLTLWSPSADKVSVVVYDKNDPDKVVGTVALEKGERGTWKQTLDSTNKLGITDFTGYYYQYQIERQGKTVLALDPYAKSLAAWNSDDSKIDDAHKVAKAAFVDPAKLGPQDLTYGKIHNFKTREDAVIYEAHVRDFTSDPAIAKDLTKPFGTFEAFIEKLDYLKDLGVTHIQLLPVLSYYFVNELKNHERLSDYASSNSNYNWGYDPQNYFSLTGMYSSDPKNPEKRIAEFKNLINEIHKRGMGAILDVVYNHTAKVDLFEDLEPNYYHFMDADGTPRTSFGGGRLGTTHHMTKRLLIDSIKYLVDTYKVDGFRFDMMGDHDAASIEEAYKAARALNPNLIMLGEGWRTYAGDENMPTKAADQDWMKHTDTVAVFSDDIRNNLKSGYPNEGQPAFITGGKRDVNTIFKNLIAQPTNFEADSPGDVIQYIAAHDNLTLFDIIAQSIKKDPSKAENYAEIHRRLRLGNLMVLTAQGTPFIHSGQEYGRTKQFRDPAYKTPVAEDKVPNKSHLLRDKDGNPFDYPYFIHDSYDSSDAVNKFDWTKATDGKAYPENVKSRDYMKGLIALRQSTDAFRLKSLQDIKDRVHLITVPGQNGVEKEDVVIGYQITAPNGDIYAVFVNADEKAREFNLGTAFAHLRNAEVLADENQAGPVGIANPKGLEWTEKGLKLNALTATVLRVSQNGTSHESTAEEKPDSTPSKPEHQNEASHPAHQDPAPEARPDSTKPDAKVADAENKPSQATADSQAEQPAQEAQASSVKEAVRNESVENSSKENIPATPDKQAELPNTGIKNENKLLFAGISLLALLGLGFLLKNKKEN |
| Enzyme Length | 1280 |
| Uniprot Accession Number | A0A0H2UNG0 |
| Absorption | |
| Active Site | ACT_SITE 778; /note=Nucleophile; /evidence=ECO:0000269|PubMed:21565699; ACT_SITE 807; /note=Proton donor; /evidence=ECO:0000269|PubMed:21565699 |
| Activity Regulation | ACTIVITY REGULATION: Inhibited by 4-O-alpha-D-glucopyranosylmoranoline (G1M). {ECO:0000269|PubMed:21565699}. |
| Binding Site | BINDING 168; /note="Substrate"; /evidence="ECO:0000269|PubMed:17187076, ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2J44, ECO:0007744|PDB:2YA1"; BINDING 214; /note="Substrate"; /evidence="ECO:0000269|PubMed:17187076, ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2J44, ECO:0007744|PDB:2YA1"; BINDING 276; /note="Substrate"; /evidence="ECO:0000269|PubMed:17187076, ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2J44, ECO:0007744|PDB:2YA1"; BINDING 318; /note="Substrate"; /evidence="ECO:0000269|PubMed:17187076, ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2J44, ECO:0007744|PDB:2YA1"; BINDING 323; /note="Substrate"; /evidence="ECO:0000269|PubMed:17187076, ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2J44, ECO:0007744|PDB:2YA1"; BINDING 743; /note="Substrate; via amide nitrogen"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA1"; BINDING 809; /note="Substrate"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA1"; BINDING 839; /note="Substrate"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA1"; BINDING 842; /note="Substrate"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA1"; BINDING 849; /note="Substrate"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA1"; BINDING 896; /note="Substrate"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA1"; BINDING 969; /note="Substrate"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA1" |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=Hydrolysis of (1->6)-alpha-D-glucosidic linkages in pullulan, amylopectin and glycogen, and in the alpha- and beta-limit dextrins of amylopectin and glycogen.; EC=3.2.1.41; Evidence={ECO:0000269|PubMed:20497336, ECO:0000269|PubMed:21565699}; |
| DNA Binding | |
| EC Number | 3.2.1.41 |
| Enzyme Function | FUNCTION: Virulence factor (By similarity). Involved in the degradation of glycogen of the mammalian host cells (PubMed:21565699). Hydrolyzes the alpha-1,6-branchpoints of glycogen (PubMed:20497336, PubMed:21565699). Hydrolyzes pullulan. Does not hydrolyze dextran (PubMed:20497336). Binds to mouse lung alveolar type II cells that are rich in glycogen stores. Is an alpha-glucan-specific carbohydrate-binding protein, which binds to amylose (pure alpha-(1,4)-linked glucose), amylopectin (alpha-(1,4)-linked glucose with alpha-(1,6) branch points), pullulan (linear polymer of mixed alpha-(1,4)- and alpha-(1,6)-linked glucose) and glycogen (similar to amylopectin with more frequent alpha-(1,6) branch points) in vitro. Does not bind to dextran (a linear polymer of alpha-(1,6)-linked glucose) (PubMed:17187076). {ECO:0000250|UniProtKB:A0A0H2ZL64, ECO:0000269|PubMed:17187076, ECO:0000269|PubMed:20497336, ECO:0000269|PubMed:21565699}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (2); Beta strand (63); Binding site (12); Chain (1); Compositional bias (4); Helix (34); Metal binding (8); Modified residue (1); Motif (1); Mutagenesis (3); Propeptide (1); Region (6); Signal peptide (1); Site (1); Turn (10) |
| Keywords | 3D-structure;Calcium;Cell wall;Glycosidase;Hydrolase;Metal-binding;Peptidoglycan-anchor;Secreted;Signal;Virulence |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: Secreted, cell wall {ECO:0000255|PROSITE-ProRule:PRU00477}; Peptidoglycan-anchor {ECO:0000255|PROSITE-ProRule:PRU00477}. Cell surface {ECO:0000250|UniProtKB:Q9F930}. Note=Localizes to cytoplasm in the lung alveolar type II cells of the mouse and human hosts. {ECO:0000269|PubMed:17187076, ECO:0000269|PubMed:21565699}. |
| Modified Residue | MOD_RES 1249; /note=Pentaglycyl murein peptidoglycan amidated threonine; /evidence=ECO:0000255|PROSITE-ProRule:PRU00477 |
| Post Translational Modification | |
| Signal Peptide | SIGNAL 1..44; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (4) |
| Cross Reference PDB | 2J44; 2YA0; 2YA1; 2YA2; |
| Mapped Pubmed ID | - |
| Motif | MOTIF 1246..1250; /note=LPXTG sorting signal; /evidence=ECO:0000255|PROSITE-ProRule:PRU00477 |
| Gene Encoded By | |
| Mass | 142,619 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=360 uM for para-nitrophenyl-alpha-D-maltopentaoside (pNP-M5) (at 25 degrees Celsius) {ECO:0000269|PubMed:21565699}; Note=kcat is 270 min(-1) for pNP-M5. {ECO:0000269|PubMed:21565699}; |
| Metal Binding | METAL 661; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA0"; METAL 663; /note="Calcium 1; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA0"; METAL 828; /note="Calcium 2; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA2"; METAL 831; /note="Calcium 2; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA2"; METAL 832; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA2"; METAL 882; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA2"; METAL 886; /note="Calcium 2"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA2"; METAL 992; /note="Calcium 1"; /evidence="ECO:0000269|PubMed:21565699, ECO:0007744|PDB:2YA0" |
| Rhea ID | |
| Cross Reference Brenda |