CHS_ECOLX
ID CHS_ECOLX Reviewed; 686 AA.
AC Q8L0V4;
DT 26-APR-2005, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2002, sequence version 1.
DT 03-AUG-2022, entry version 64.
DE RecName: Full=Chondroitin synthase;
DE Short=CS;
DE AltName: Full=Chondroitin polymerase;
DE Includes:
DE RecName: Full=Glucuronosyl-N-acetylgalactosaminyl-proteoglycan 4-beta-N-acetylgalactosaminyltransferase;
DE EC=2.4.1.175 {ECO:0000269|PubMed:11943778};
DE AltName: Full=UDP-GalNAc transferase;
DE Includes:
DE RecName: Full=N-acetylgalactosaminyl-proteoglycan 3-beta-glucuronosyltransferase;
DE EC=2.4.1.226 {ECO:0000269|PubMed:11943778};
DE AltName: Full=UDP-GlcUA transferase;
GN Name=kfoC;
OS Escherichia coli.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=O5:K4(L):H4 / ATCC 23502;
RX PubMed=11943778; DOI=10.1074/jbc.m201719200;
RA Ninomiya T., Sugiura N., Tawada A., Sugimoto K., Watanabe H., Kimata K.;
RT "Molecular cloning and characterization of chondroitin polymerase from
RT Escherichia coli strain K4.";
RL J. Biol. Chem. 277:21567-21575(2002).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) OF 58-682 IN COMPLEXES WITH UDP;
RP UDP-ALPHA-D-GLUCURONATE AND UDP-N-ACETYL-ALPHA-D-GALACTOSAMINE, COFACTOR,
RP AND SUBSTRATE SPECIFICITY.
RC STRAIN=O5:K4(L):H4 / ATCC 23502;
RX PubMed=18771653; DOI=10.1016/j.bbrc.2008.08.121;
RA Osawa T., Sugiura N., Shimada H., Hirooka R., Tsuji A., Shirakawa T.,
RA Fukuyama K., Kimura M., Kimata K., Kakuta Y.;
RT "Crystal structure of chondroitin polymerase from Escherichia coli K4.";
RL Biochem. Biophys. Res. Commun. 378:10-14(2009).
CC -!- FUNCTION: Glycosyltransferase that catalyzes elongation of chondroitin,
CC a polysaccharide composed of a repeating disaccharide of N-
CC acetylgalactosamine (GalNAc) and glucuronic acid (GlcUA) units, by
CC alternatively transferring the GlcUA and GalNAc moiety from UDP-GlcUA
CC and UDP-GalNAc to the non-reducing ends of the chondroitin chain. Each
CC chondroitin unit has the composition beta-(1->4)-GlcUA-beta-(1->3)-
CC GalNAc. {ECO:0000269|PubMed:11943778}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-O-(beta-D-GlcA-(1->3)-beta-D-GalNAc-(1->4)-beta-D-GlcA-
CC (1->3)-beta-D-Gal-(1->3)-beta-D-Gal-(1->4)-beta-D-Xyl)-L-seryl-
CC [protein] + UDP-N-acetyl-alpha-D-galactosamine = 3-O-(beta-D-GalNAc-
CC (1->4)-beta-D-GlcA-(1->3)-beta-D-GalNAc-(1->4)-beta-D-GlcA-(1->3)-
CC beta-D-Gal-(1->3)-beta-D-Gal-(1->4)-beta-D-Xyl)-L-seryl-[protein] +
CC H(+) + UDP; Xref=Rhea:RHEA:20800, Rhea:RHEA-COMP:14058, Rhea:RHEA-
CC COMP:14059, ChEBI:CHEBI:15378, ChEBI:CHEBI:58223, ChEBI:CHEBI:67138,
CC ChEBI:CHEBI:138442, ChEBI:CHEBI:138443; EC=2.4.1.175;
CC Evidence={ECO:0000269|PubMed:11943778};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-O-(beta-D-GlcA-(1->3)-[beta-D-GalNAc-(1->4)-beta-D-GlcA-
CC (1->3)](n)-beta-D-GalNAc-(1->4)-beta-D-GlcA-(1->3)-beta-D-Gal-(1->3)-
CC beta-D-Gal-(1->4)-beta-D-Xyl)-L-seryl-[protein] + UDP-N-acetyl-alpha-
CC D-galactosamine = 3-O-([beta-D-GalNAc-(1->4)-beta-D-GlcA-
CC (1->3)](n+1)-beta-D-GalNAc-(1->4)-beta-D-GlcA-(1->3)-beta-D-Gal-
CC (1->3)-beta-D-Gal-(1->4)-beta-D-Xyl)-L-seryl-[protein] + H(+) + UDP;
CC Xref=Rhea:RHEA:55000, Rhea:RHEA-COMP:14060, Rhea:RHEA-COMP:14301,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:58223, ChEBI:CHEBI:67138,
CC ChEBI:CHEBI:138444, ChEBI:CHEBI:138445; EC=2.4.1.175;
CC Evidence={ECO:0000269|PubMed:11943778};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-O-(beta-D-GalNAc-(1->4)-beta-D-GlcA-(1->3)-beta-D-Gal-
CC (1->3)-beta-D-Gal-(1->4)-beta-D-Xyl)-L-seryl-[protein] + UDP-alpha-D-
CC glucuronate = 3-O-(beta-D-GlcA-(1->3)-beta-D-GalNAc-(1->4)-beta-D-
CC GlcA-(1->3)-beta-D-Gal-(1->3)-beta-D-Gal-(1->4)-beta-D-Xyl)-L-seryl-
CC [protein] + H(+) + UDP; Xref=Rhea:RHEA:23428, Rhea:RHEA-COMP:12575,
CC Rhea:RHEA-COMP:14058, ChEBI:CHEBI:15378, ChEBI:CHEBI:58052,
CC ChEBI:CHEBI:58223, ChEBI:CHEBI:132105, ChEBI:CHEBI:138442;
CC EC=2.4.1.226; Evidence={ECO:0000269|PubMed:11943778};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-O-([beta-D-GalNAc-(1->4)-beta-D-GlcA-(1->3)](n)-beta-D-
CC GalNAc-(1->4)-beta-D-GlcA-(1->3)-beta-D-Gal-(1->3)-beta-D-Gal-(1->4)-
CC beta-D-Xyl)-L-seryl-[protein] + UDP-alpha-D-glucuronate = 3-O-(beta-
CC D-GlcA-(1->3)-[beta-D-GalNAc-(1->4)-beta-D-GlcA-(1->3)](n)-beta-D-
CC GalNAc-(1->4)-beta-D-GlcA-(1->3)-beta-D-Gal-(1->3)-beta-D-Gal-(1->4)-
CC beta-D-Xyl)-L-seryl-[protein] + H(+) + UDP; Xref=Rhea:RHEA:54996,
CC Rhea:RHEA-COMP:14060, Rhea:RHEA-COMP:14061, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:58052, ChEBI:CHEBI:58223, ChEBI:CHEBI:138444,
CC ChEBI:CHEBI:138445; EC=2.4.1.226;
CC Evidence={ECO:0000269|PubMed:11943778};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:11943778, ECO:0000269|PubMed:18771653};
CC Note=Binds 2 Mn(2+) ions per subunit. {ECO:0000269|PubMed:18771653};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.44 uM for UDP-GlcUA {ECO:0000269|PubMed:11943778};
CC KM=31.6 uM for UDP-GalNAc {ECO:0000269|PubMed:11943778};
CC pH dependence:
CC Optimum pH is 7-7.5. {ECO:0000269|PubMed:11943778};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius for highest reaction speed,
CC and 25 degrees Celsius to obtain highest molecular weight of product
CC chondroitin.;
CC -!- SIMILARITY: Belongs to the glycosyltransferase 2 family. CS/HAS
CC subfamily. {ECO:0000305}.
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DR EMBL; AB079602; BAC00523.1; -; Genomic_DNA.
DR RefSeq; WP_000025667.1; NZ_WSWP01000006.1.
DR PDB; 2Z86; X-ray; 2.40 A; A/B/C/D=58-682.
DR PDB; 2Z87; X-ray; 3.00 A; A/B=59-682.
DR PDBsum; 2Z86; -.
DR PDBsum; 2Z87; -.
DR AlphaFoldDB; Q8L0V4; -.
DR SMR; Q8L0V4; -.
DR CAZy; GT2; Glycosyltransferase Family 2.
DR KEGG; ag:BAC00523; -.
DR BRENDA; 2.4.1.175; 2026.
DR EvolutionaryTrace; Q8L0V4; -.
DR GO; GO:0047238; F:glucuronosyl-N-acetylgalactosaminyl-proteoglycan 4-beta-N-acetylgalactosaminyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0050510; F:N-acetylgalactosaminyl-proteoglycan 3-beta-glucuronosyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0008152; P:metabolic process; IEA:UniProtKB-KW.
DR Gene3D; 3.90.550.10; -; 2.
DR InterPro; IPR001173; Glyco_trans_2-like.
DR InterPro; IPR029044; Nucleotide-diphossugar_trans.
DR Pfam; PF00535; Glycos_transf_2; 2.
DR SUPFAM; SSF53448; SSF53448; 2.
PE 1: Evidence at protein level;
KW 3D-structure; Glycosyltransferase; Manganese; Metal-binding;
KW Multifunctional enzyme; Repeat; Transferase.
FT CHAIN 1..686
FT /note="Chondroitin synthase"
FT /id="PRO_0000059257"
FT REGION 130..417
FT /note="Galactosaminyltransferase; A1 domain"
FT /evidence="ECO:0000269|PubMed:18771653"
FT REGION 418..682
FT /note="Glucuronosyltransferase; A2 domain"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 157
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 161
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 188
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 217
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 223
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 239..240
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 241
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 361..362
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 386
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 441
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 469
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 517..520
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 521
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 581
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 603..604
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT /evidence="ECO:0000269|PubMed:18771653"
FT BINDING 631
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:18771653"
FT HELIX 65..73
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 75..77
FT /evidence="ECO:0007829|PDB:2Z87"
FT HELIX 81..94
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 130..134
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 152..160
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 162..173
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 181..189
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 194..199
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 200..204
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 207..212
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 219..229
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 232..238
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 242..244
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 248..258
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 262..265
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 268..271
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 273..275
FT /evidence="ECO:0007829|PDB:2Z87"
FT HELIX 278..283
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 287..289
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 300..302
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 303..307
FT /evidence="ECO:0007829|PDB:2Z87"
FT HELIX 313..319
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 320..325
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 329..332
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 337..341
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 343..348
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 361..371
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 375..378
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 380..382
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 384..387
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 391..393
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 408..412
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 414..417
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 423..425
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 433..442
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 444..446
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 447..455
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 456..458
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 461..470
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 472..474
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 475..483
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 489..494
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 499..509
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 512..517
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 528..538
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 543..552
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 558..561
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 570..573
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 583..586
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 587..590
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 591..594
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 604..612
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 613..615
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 618..629
FT /evidence="ECO:0007829|PDB:2Z86"
FT HELIX 640..654
FT /evidence="ECO:0007829|PDB:2Z86"
FT TURN 655..659
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 662..668
FT /evidence="ECO:0007829|PDB:2Z86"
FT STRAND 677..681
FT /evidence="ECO:0007829|PDB:2Z86"
SQ SEQUENCE 686 AA; 79257 MW; 6FB941623D9EC9D4 CRC64;
MSILNQAINL YKNKNYRQAL SLFEKVAEIY DVSWVEANIK LCQTALNLSE EVDKLNRKAV
IDIDAATKIM CSNAKAISLN EVEKNEIISK YREITAKKSE RAELKEVEPI PLDWPSDLTL
PPLPESTNDY VWAGKRKELD DYPRKQLIID GLSIVIPTYN RAKILAITLA CLCNQKTIYD
YEVIVADDGS KENIEEIVRE FESLLNIKYV RQKDYGYQLC AVRNLGLRAA KYNYVAILDC
DMAPNPLWVQ SYMELLAVDD NVALIGPRKY IDTSKHTYLD FLSQKSLINE IPEIITNNQV
AGKVEQNKSV DWRIEHFKNT DNLRLCNTPF RFFSGGNVAF AKKWLFRAGW FDEEFTHWGG
EDNEFGYRLY REGCYFRSVE GAMAYHQEPP GKENETDRAA GKNITVQLLQ QKVPYFYRKK
EKIESATLKR VPLVSIYIPA YNCSKYIVRC VESALNQTIT DLEVCICDDG STDDTLRILQ
EHYANHPRVR FISQKNKGIG SASNTAVRLC RGFYIGQLDS DDFLEPDAVE LCLDEFRKDL
SLACVYTTNR NIDREGNLIS NGYNWPIYSR EKLTSAMICH HFRMFTARAW NLTEGFNESI
SNAVDYDMYL KLSEVGPFKH INKICYNRVL HGENTSIKKL DIQKENHFKV VNESLSRLGI
KKYKYSPLTN LNECRKYTWE KIENDL
