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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)
 
 
Protein Information
Protein Name
Chondroitinase ABC
UniProtKB/SwissProt ID
UniProtKB Sequence
>sp|P59807|CABC_PROVU Chondroitin ABC endolyase 1 OS=Proteus vulgaris PE=1 SV=1
MPIFRFTALAMTLGLLSAPYNAMAATSNPAFDPKNLMQSEIYHFAQNNPLADFSSDKNSI LTLSDKRSIMGNQSL
LWKWKGGSSFTLHKKLIVPTDKEASKAWGRSSTPVFSFWLYNEKP IDGYLTIDFGEKLISTSEAQAGFKVKLDFT
GWRAVGVSLNNDLENREMTLNATNTSSDGT QDSIGRSLGAKVDSIRFKAPSNVSQGEIYIDRIMFSVDDARYQWS
DYQVKTRLSEPEIQF HNVKPQLPVTPENLAAIDLIRQRLINEFVGGEKETNLALEENISKLKSDFDALNIHTLAN
GGTQGRHLITDKQIIIYQPENLNSQDKQLFDNYVILGNYTTLMFNISRAYVLEKDPTQKA QLKQMYLLMTKHLL
DQGFVKGSALVTTHHWGYSSRWWYISTLLMSDALKEANLQTQVYDS LLWYSREFKSSFDMKVSADSSDLDYFNTL
SRQHLALLLLEPDDQKRINLVNTFSHYITGA LTQVPPGGKDGLRPDGTAWRHEGNYPGYSFPAFKNASQLIYLLR
DTPFSVGESGWNNLKK AMVSAWIYSNPEVGLPLAGRHPFNSPSLKSVAQGYYWLAMSAKSSPDKTLASIYLAISD
K TQNESTAIFGETITPASLPQGFYAFNGGAFGIHRWQDKMVTLKAYNTNVWSSEIYNKDNR YGRYQSHGVAQIV
SNGSQLSQGYQQEGWDWNRMEGATTIHLPLKDLDSPKPHTLMQRGER GFSGTSSLEGQYGMMAFNLIYPANLERF
DPNFTAKKSVLAADNHLIFIGSNINSSDKNKN VETTLFQHAITPTLNTLWINGQKIENMPYQTTLQQGDWLIDSN
GNGYLITQAEKVNVSRQ HQVSAENKNRQPTEGNFSSAWIDHSTRPKDASYEYMVFLDATPEKMGEMAQKFRENNG
LY QVLRKDKDVHIILDKLSNVTGYAFYQPASIEDKWIKKVNKPAIVMTHRQKDTLIVSAVTP DLNMTRQKAATP
VTINVTINGKWQSADKNSEVKYQVSGDNTELTFTSYFGIPQEIKLSPL 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
 
 
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 LTLSDKRSIMGNQ
SLLWKWKGGSSFTLHKKLIVPTDKEASKAWGRSSTPVFSFWLYNEKP IDGYLTIDFGEKLISTSEAQAGFKVKL
DFTGWRAVGVSLNNDLENREMTLNATNTSSDGT QDSIGRSLGAKVDSIRFKAPSNVSQGEIYIDRIMFSVDDAR
YQWSDYQVKTRLSEPEIQF HNVKPQLPVTPENLAAIDLIRQRLINEFVGGEKETNLALEEN*(282)ISKLK
SDFDALNIHTLAN GGTQGRHLITDKQIIIYQPENLNSQDKQLFDNYVILGN*(338)YTTLMFN*(345)
ISRAYVLEKDPTQKA QLKQMYLLMTKHLLDQGFVKGSALVTTHHWGYSSRWWYISTLLMSDALKEANLQTQVYD
S LLWYSREFKSSFDMKVSADSSDLDYFNTLSRQHLALLLLEPDDQKRINLVNTFSHYITGA LTQVPPGGKDG
LRPDGTAWRHEGNYPGYSFPAFKN*(515)ASQLIYLLRDTPFSVGESGWNNLKK AMVSAWIYSNPEVGLPL
AGRHPFNSPSLKSVAQGYYWLAMSAKSSPDKTLASIYLAISDK TQNESTAIFGETITPASLPQGFYAFNGGAFG
IHRWQDKMVTLKAYNTNVWSSEIYNKDNR YGRYQSHGVAQIVSN*(675)GSQLSQGYQQEGWDWNRMEGAT
TIHLPLKDLDSPKPHTLMQRGER GFSGTSSLEGQYGMMAFNLIYPANLERFDPNFTAKKSVLAADNHLIFIGSN
INSSDKNKN VETTLFQHAITPTLNTLWINGQKIENMPYQTTLQQGDWLIDSNGNGYLITQAEKVNVSRQ HQV
SAENKNRQPTEGN*(856)FSSAWIDHSTRPKDASYEYMVFLDATPEKMGEMAQKFRENNGLY QVLRKDKDV
HIILDKLSNVTGYAFYQPASIEDKWIKKVNKPAIVMTHRQKDTLIVSAVTP DLN*(963)MTRQKAATPVTI
NVTINGKWQSADKNSEVKYQVSGDNTELTFTSYFGIPQEIKLSPL P
Sequence Around Glycosites (21 AA)
EKETNLALEENISKLKSDFDA
QLFDNYVILGNYTTLMFNISR
ILGNYTTLMFNISRAYVLEKD
YPGYSFPAFKNASQLIYLLRD
QSHGVAQIVSNGSQLSQGYQQ
ENKNRQPTEGNFSSAWIDHST
LIVSAVTPDLNMTRQKAATPV
Glycosite Sequence Logo
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.
 
 
Literature
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
 
 
Copyright @ 2011 IMTECH
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