ProGlyProt ID BC165 Organism Information Organism Name Proteus vulgaris Domain Bacteria Classification Family: Enterobacteriaceae Order: "Enterobacteriales" Class: Gammaproteobacteria Division or phylum: "Proteobacteria" Taxonomic ID (NCBI) 585 Genome Sequence (s) EMBL Gene Information Gene Name NCBI Gene ID Protein Information Protein Name Chondroitinase ABC UniProtKB/SwissProt ID P59807 NCBI RefSeq EMBL-CDS UniProtKB Sequence >sp|P59807|CABC_PROVU Chondroitin ABC endolyase 1 OS=Proteus vulgaris PE=1 SV=1 MPIFRFTALAMTLGLLSAPYNAMAATSNPAFDPKNLMQSEIYHFAQNNPLADFSSDKNSI LTLSDKRSIMGNQSLLWKWKGGSSFTLHKKLIVPTDKEASKAWGRSSTPVFSFWLYNEKP IDGYLTIDFGEKLISTSEAQAGFKVKLDFTGWRAVGVSLNNDLENREMTLNATNTSSDGT QDSIGRSLGAKVDSIRFKAPSNVSQGEIYIDRIMFSVDDARYQWSDYQVKTRLSEPEIQF HNVKPQLPVTPENLAAIDLIRQRLINEFVGGEKETNLALEENISKLKSDFDALNIHTLAN GGTQGRHLITDKQIIIYQPENLNSQDKQLFDNYVILGNYTTLMFNISRAYVLEKDPTQKA QLKQMYLLMTKHLLDQGFVKGSALVTTHHWGYSSRWWYISTLLMSDALKEANLQTQVYDS LLWYSREFKSSFDMKVSADSSDLDYFNTLSRQHLALLLLEPDDQKRINLVNTFSHYITGA LTQVPPGGKDGLRPDGTAWRHEGNYPGYSFPAFKNASQLIYLLRDTPFSVGESGWNNLKK AMVSAWIYSNPEVGLPLAGRHPFNSPSLKSVAQGYYWLAMSAKSSPDKTLASIYLAISDK TQNESTAIFGETITPASLPQGFYAFNGGAFGIHRWQDKMVTLKAYNTNVWSSEIYNKDNR YGRYQSHGVAQIVSNGSQLSQGYQQEGWDWNRMEGATTIHLPLKDLDSPKPHTLMQRGER GFSGTSSLEGQYGMMAFNLIYPANLERFDPNFTAKKSVLAADNHLIFIGSNINSSDKNKN VETTLFQHAITPTLNTLWINGQKIENMPYQTTLQQGDWLIDSNGNGYLITQAEKVNVSRQ HQVSAENKNRQPTEGNFSSAWIDHSTRPKDASYEYMVFLDATPEKMGEMAQKFRENNGLY QVLRKDKDVHIILDKLSNVTGYAFYQPASIEDKWIKKVNKPAIVMTHRQKDTLIVSAVTP DLNMTRQKAATPVTINVTINGKWQSADKNSEVKYQVSGDNTELTFTSYFGIPQEIKLSPL P Sequence length 1021 AA Subcellular Location Secreted Function Chondroitin ABC endolyase 1 is a bacterial enzyme that degrades long sulphated glycosaminoglycan (GAG) chains of chondroitin sulphate proteoglycans (CSPGs). CSPGs are responsible for axon growth inhibition. EC: 4.2.2.20. Protein Structure PDB ID 1HN0 Glycosylation Status Glycosylation Type N (Asn) linked Experimentally Validated Glycosite(s) in Full Length Protein N282, N338, N345, N515, N675, N856, N963 Experimentally Validated Glycosite(s ) in Mature Protein N282, N338, N345, N515, N675, N856, N963 Glycosite(s) Annotated Protein Sequence >sp|P59807|CABC_PROVU Chondroitin ABC endolyase 1 OS=Proteus vulgaris PE=1 SV=1 MPIFRFTALAMTLGLLSAPYNAMAATSNPAFDPKNLMQSEIYHFAQNNPLADFSSDKNSI LTLSDKRSIMGNQSLLWKWKGGSSFTLHKKLIVPTDKEASKAWGRSSTPVFSFWLYNEKP IDGYLTIDFGEKLISTSEAQAGFKVKLDFTGWRAVGVSLNNDLENREMTLNATNTSSDGT QDSIGRSLGAKVDSIRFKAPSNVSQGEIYIDRIMFSVDDARYQWSDYQVKTRLSEPEIQF HNVKPQLPVTPENLAAIDLIRQRLINEFVGGEKETNLALEEN*(282)ISKLKSDFDALNIHTLAN GGTQGRHLITDKQIIIYQPENLNSQDKQLFDNYVILGN*(338)YTTLMFN*(345)ISRAYVLEKDPTQKA QLKQMYLLMTKHLLDQGFVKGSALVTTHHWGYSSRWWYISTLLMSDALKEANLQTQVYDS LLWYSREFKSSFDMKVSADSSDLDYFNTLSRQHLALLLLEPDDQKRINLVNTFSHYITGA LTQVPPGGKDGLRPDGTAWRHEGNYPGYSFPAFKN*(515)ASQLIYLLRDTPFSVGESGWNNLKK AMVSAWIYSNPEVGLPLAGRHPFNSPSLKSVAQGYYWLAMSAKSSPDKTLASIYLAISDK TQNESTAIFGETITPASLPQGFYAFNGGAFGIHRWQDKMVTLKAYNTNVWSSEIYNKDNR YGRYQSHGVAQIVSN*(675)GSQLSQGYQQEGWDWNRMEGATTIHLPLKDLDSPKPHTLMQRGER GFSGTSSLEGQYGMMAFNLIYPANLERFDPNFTAKKSVLAADNHLIFIGSNINSSDKNKN VETTLFQHAITPTLNTLWINGQKIENMPYQTTLQQGDWLIDSNGNGYLITQAEKVNVSRQ HQVSAENKNRQPTEGN*(856)FSSAWIDHSTRPKDASYEYMVFLDATPEKMGEMAQKFRENNGLY QVLRKDKDVHIILDKLSNVTGYAFYQPASIEDKWIKKVNKPAIVMTHRQKDTLIVSAVTP DLN*(963)MTRQKAATPVTINVTINGKWQSADKNSEVKYQVSGDNTELTFTSYFGIPQEIKLSPL P Sequence Around Glycosites (21 AA) EKETNLALEENISKLKSDFDA QLFDNYVILGNYTTLMFNISR ILGNYTTLMFNISRAYVLEKD YPGYSFPAFKNASQLIYLLRD QSHGVAQIVSNGSQLSQGYQQ ENKNRQPTEGNFSSAWIDHST LIVSAVTPDLNMTRQKAATPV Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Slower migration on SDS-PAGE than its predicted mass and N-glycosidase treatment Technique(s) used for Glycosylated Residue(s) Detection Multi-site-directed mutagenesis (mostly Asn→Gln) Protein Glycosylation- Implication Carbohydrate chains either distort the protein structure or produce steric hindrance of the active site resulting in an inactive form of the secreted protein. Secretion of underglycosylated protein was better when expressed in mammalian cells compared to fully glycosylated protein. Glycan Information Glycan Annotation Technique(s) used for Glycan Identification Protein Glycosylation linked (PGL) gene(s) OST Gene Name OST NCBI Gene ID OST GenBank Gene Sequence OST Protein Name OST UniProtKB/ SwissProt ID OST NCBI RefSeq OST EMBL-CDS OST UniProtKB Sequence OST EC Number (BRENDA) OST Genome Context Characterized Accessory Gene(s) PGL Additional Links CAZy Literatures Reference(s) 1) Muir, E.M., Fyfe, I., Gardiner, S., Li, L., Warren, P., Fawcett, J.W., Keynes, R.J. and Rogers, J.H. (2010) Modification of N-glycosylation sites allows secretion of bacterial chondroitinase ABC from mammalian cells. J Biotechnol, 145, 103-110. [PubMed: 19900493] 2) Huang, W., Lunin, V.V., Li, Y., Suzuki, S., Sugiura, N., Miyazono, H. and Cygler, M. (2003) Crystal structure of Proteus vulgaris chondroitin sulfate ABC lyase I at 1.9A resolution. J Mol Biol, 328, 623-634. [PubMed: 12706721 Additional Comments Engineered glycoprotein Sequon features: eukaryotic consensus sequence NXS/T gets glycosylated. Chondroitinase ABC of Proteus vulgaris is an example of a protein of bacterial origin that is expressed as glycoprotein in mammalian cells. The glycosylated residues were mutated to abolish glycosylation that interferes with protein folding and secretion. Year of Identification 2010 Year of Validation 2010