EzCatDB: T00415
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DB codeT00415
RLCP classification3.903.70210.640 : Transfer
CATH domainDomain 1-.-.-.-
Domain 23.90.550.10 : Spore Coat Polysaccharide Biosynthesis Protein SpsA; Chain ACatalytic domain
Domain 33.10.180.20 : 2,3-Dihydroxybiphenyl 1,2-Dioxygenase; domain 1
E.C.2.4.1.101
CSA1foa

CATH domainRelated DB codes (homologues)
3.90.550.10 : Spore Coat Polysaccharide Biosynthesis Protein SpsA; Chain AS00709,S00465,S00466,D00417,D00859,D00860

Enzyme Name
UniProtKBKEGG

P27115
Protein nameAlpha-1,3-mannosyl-glycoprotein 2-beta-N-acetylglucosaminyltransferaseAlpha-1,3-mannosyl-glycoprotein 2-beta-N-acetylglucosaminyltransferase
N-Acetylglucosaminyltransferase I
N-Glycosyl-oligosaccharide-glycoprotein N-acetylglucosaminyltransferase I
Uridine diphosphoacetylglucosamine-alpha-1,3-mannosylglycoprotein beta-1,2-N-acetylglucosaminyltransferase
UDP-N-acetylglucosaminyl:alpha-1,3-D-mannoside-beta-1,2-N-acetylglucosaminyltransferase I
UDP-N-acetylglucosaminyl:alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I
Alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase
GnTI
SynonymsEC 2.4.1.101
N-glycosyl-oligosaccharide-glycoprotein N-acetylglucosaminyltransferase I
GNT-I
GlcNAc-T I
RefSeqNP_001076214.1 (Protein)
NM_001082745.1 (DNA/RNA sequence)
PfamPF03071 (GNT-I)
[Graphical view]
CAZyGT13 (Glycosyltransferase Family)

KEGG pathways
MAP codePathways
MAP00510N-Glycan biosynthesis
MAP01030Glycan structures - biosynthesis 1

UniProtKB:Accession NumberP27115
Entry nameMGAT1_RABIT
ActivityUDP-N-acetyl-D-glucosamine + 3-(alpha-D-mannosyl)-beta-D-mannosyl-R = UDP + 3-(2-(N-acetyl-beta-D-glucosaminyl)-alpha-D-mannosyl)-beta-D-mannosyl-R.
Subunit
Subcellular locationGolgi apparatus membrane, Single-pass type II membrane protein.
Cofactor

Compound table: links to PDB-related databases & PoSSuM

CofactorsSubstratesProducts
KEGG-idC00034C00043C04614C00015C04880
CompoundManganeseUDP-N-acetyl-D-glucosamine3-(alpha-D-mannosyl)-beta-D-mannosyl-RUDP3-(2-[N-acetyl-beta-D-glucosaminyl]-alpha-D-mannosyl)-beta-D-mannosyl-R
Typeheavy metalamide group,carbohydrate,nucleotidepolysaccharideamide group,nucleotideamide group,polysaccharide
ChEBI18291
35154
16264

17659

PubChem23930
445675

6031

             
1fo8A01UnboundUnboundUnboundUnboundUnbound
1fo9A01UnboundUnboundUnboundUnboundUnbound
1foaA01Bound:_MNBound:UD1UnboundUnboundUnbound
2am3A01Bound:_MNAnalogue:UPGAnalogue:GOL 450UnboundUnbound
2am4A01Bound:_MNAnalogue:U2FUnboundUnboundUnbound
2am5A01Bound:_MNUnboundUnboundBound:UDPUnbound
2apcA01UnboundAnalogue:UDMAnalogue:GOL 450UnboundUnbound
1fo8A02UnboundUnboundUnboundUnboundUnbound
1fo9A02UnboundUnboundUnboundUnboundUnbound
1foaA02UnboundUnboundUnboundUnboundUnbound
2am3A02UnboundUnboundUnboundUnboundUnbound
2am4A02UnboundUnboundUnboundUnboundUnbound
2am5A02UnboundUnboundUnboundUnboundUnbound
2apcA02UnboundUnboundUnboundUnboundUnbound

Active-site residues
resource
literature [1], [3], [6], [7], [8]
pdbCatalytic residuesCofactor-binding residues
          
1fo8A01ASP 291
ASP 213(Manganese binding)
1fo9A01ASP 291
ASP 213(Manganese binding)
1foaA01ASP 291
ASP 213(Manganese binding)
2am3A01ASP 291
ASP 213(Manganese binding)
2am4A01ASP 291
ASP 213(Manganese binding)
2am5A01ASP 291
ASP 213(Manganese binding)
2apcA01ASP 291
ASP 213(Manganese binding)
1fo8A02 
 
1fo9A02 
 
1foaA02 
 
2am3A02 
 
2am4A02 
 
2am5A02 
 
2apcA02 
 

References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[1]Fig.1, p.6383-6385
[3]Fig.4, p.5272-5273
[6]Fig.5, p.16924-16937
[7]p.74-76
[8]Fig.4, p.527-530

references
[1]
CommentsX-RAY CRYSTALLOGRAPHY (1.5 ANGSTROMS).
PubMed ID10350455
JournalBiochemistry
Year1999
Volume38
Pages6380-5
AuthorsCharnock SJ, Davies GJ
TitleStructure of the nucleotide-diphospho-sugar transferase, SpsA from Bacillus subtilis, in native and nucleotide-complexed forms.
Related PDB1qg8,2qg8,3qg8
Related UniProtKBP39621
[2]
PubMed ID10508766
JournalCurr Opin Struct Biol
Year1999
Volume9
Pages563-71
AuthorsBreton C, Imberty A
TitleStructure/function studies of glycosyltransferases.
[3]
PubMed ID11032794
JournalEMBO J
Year2000
Volume19
Pages5269-80
AuthorsUnligil UM, Zhou S, Yuwaraj S, Sarkar M, Schachter H, Rini JM
TitleX-ray crystal structure of rabbit N-acetylglucosaminyltransferase I: catalytic mechanism and a new protein superfamily.
Related PDB1fo8,1fo9,1foa
[4]
PubMed ID12691742
JournalJ Mol Biol
Year2003
Volume328
Pages307-17
AuthorsCoutinho PM, Deleury E, Davies GJ, Henrissat B
TitleAn evolving hierarchical family classification for glycosyltransferases.
[5]
PubMed ID15653326
JournalTrends Biochem Sci
Year2005
Volume30
Pages53-62
AuthorsQasba PK, Ramakrishnan B, Boeggeman E
TitleSubstrate-induced conformational changes in glycosyltransferases.
[6]
PubMed ID17177443
JournalJ Am Chem Soc
Year2006
Volume128
Pages16921-7
AuthorsKozmon S, Tvaroska I
TitleCatalytic mechanism of glycosyltransferases: hybrid quantum mechanical/molecular mechanical study of the inverting N-acetylglucosaminyltransferase I.
[7]
PubMed ID16769084
JournalJ Mol Biol
Year2006
Volume360
Pages67-79
AuthorsGordon RD, Sivarajah P, Satkunarajah M, Ma D, Tarling CA, Vizitiu D, Withers SG, Rini JM
TitleX-ray crystal structures of rabbit N-acetylglucosaminyltransferase I (GnT I) in complex with donor substrate analogues.
Related PDB2am3,2am4,2am5,2apc
[8]
PubMed ID18518825
JournalAnnu Rev Biochem
Year2008
Volume77
Pages521-55
AuthorsLairson LL, Henrissat B, Davies GJ, Withers SG
TitleGlycosyltransferases: structures, functions, and mechanisms.

comments
This enzyme belongs to glycosyltransferase family-13 (GT13 family), with a GT-A fold and an inverting mechanism (see [8]).
The structure of the N-terminal domain, which contains the N-terminal cytoplasmic region and a membrane-bound region, has not been determined yet.
According to the literature [3], [7] and [8], this enzyme catalyzes the following reaction:
(0) Manganese ion, bound to Asp213, stabilizes the negative charge on the leaving phosphate groups.
(1) Asp291 acts as a general base to deprotonate O2 atom of the acceptor, mannosyl group, to activate it.
(2) The activated O2 atom of the mannosyl group makes a nucleophilic attack on the C1 atom of N-acetyl-glucosamine of UDP-N-acetyl-D-glucosamine, leading to the transition state, where a positive charge is developed on the C1 atom and negative charges on the O2 atom and the leaving phosphate oxygen.
(3) The bond between the C1 atom and the leaving phosphate group is cleaved while the bond between the O2 atom and the C1 atom is formed. (SN2-like reaction)

createdupdated
2010-01-292011-10-04


Copyright: Nozomi Nagano, JST & CBRC-AIST
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Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
Funded by Grant-in-Aid for Scientific Research (B)/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (September 2005 - September 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (October 2007 - September 2010)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2011 - March 2012)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2012 - March 2013)
Supported by the commission for the Development of Artificial Gene Synthesis Technology for Creating Innovative Biomaterial from the Ministry of Economy, Trade and Industry (METI) (October 2012 - )
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