MGS_RHOMR
ID MGS_RHOMR Reviewed; 397 AA.
AC Q9RFR0;
DT 09-JUL-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2000, sequence version 1.
DT 03-AUG-2022, entry version 92.
DE RecName: Full=Mannosylglycerate synthase;
DE Short=MGS;
DE EC=2.4.1.269 {ECO:0000269|PubMed:10585410, ECO:0000269|PubMed:15951819, ECO:0000269|PubMed:21288903};
GN Name=mgs;
OS Rhodothermus marinus (Rhodothermus obamensis).
OC Bacteria; Bacteroidetes; Bacteroidetes Order II. Incertae sedis;
OC Rhodothermaceae; Rhodothermus.
OX NCBI_TaxID=29549;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-30, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBSTRATE
RP SPECIFICITY.
RC STRAIN=ATCC 43812 / R-10 / DSM 4252;
RX PubMed=10585410; DOI=10.1074/jbc.274.50.35407;
RA Martins L.O., Empadinhas N., Marugg J.D., Miguel C., Ferreira C.,
RA da Costa M.S., Santos H.;
RT "Biosynthesis of mannosylglycerate in the thermophilic bacterium
RT Rhodothermus marinus. Biochemical and genetic characterization of a
RT mannosylglycerate synthase.";
RL J. Biol. Chem. 274:35407-35414(1999).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS
RP AND DIVALENT CATIONS, FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF GLU-11;
RP ASP-100 AND ARG-131, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY,
RP AND SUBUNIT.
RX PubMed=15951819; DOI=10.1038/nsmb950;
RA Flint J., Taylor E., Yang M., Bolam D.N., Tailford L.E.,
RA Martinez-Fleites C., Dodson E.J., Davis B.G., Gilbert H.J., Davies G.J.;
RT "Structural dissection and high-throughput screening of mannosylglycerate
RT synthase.";
RL Nat. Struct. Mol. Biol. 12:608-614(2005).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) OF 1-382 IN COMPLEX WITH SUBSTRATE
RP ANALOGS, FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF LYS-9; TYR-37;
RP GLN-66; LYS-76; ASP-102; ASP-135; THR-139; TRP-189; ASP-192; HIS-217;
RP ARG-218; TYR-220 AND MET-229, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, COFACTOR, AND SUBUNIT.
RX PubMed=21288903; DOI=10.1074/jbc.m110.199844;
RA Nielsen M.M., Suits M.D., Yang M., Barry C.S., Martinez-Fleites C.,
RA Tailford L.E., Flint J.E., Dumon C., Davis B.G., Gilbert H.J., Davies G.J.;
RT "Substrate and metal ion promiscuity in mannosylglycerate synthase.";
RL J. Biol. Chem. 286:15155-15164(2011).
CC -!- FUNCTION: Involved in the biosynthesis of the stress protectant 2-O-
CC alpha-D-mannosyl glycerate (MG) which is produced in response to growth
CC at supraoptimal temperature and salinity, and protects several enzymes
CC against inactivation by temperature, freeze-drying and osmotic stress.
CC Catalyzes the condensation of alpha-GDP-D-mannose (GDP-Man) with D-
CC glycerate to produce alpha-mannosyl-D-glycerate. It is specific for
CC GDP-Man, but it can also use alpha-GDP-D-glucose (GDP-Glc), beta-GDP-D-
CC fructose, alpha-UDP-D-mannose and alpha-UDP-D-glucose as sugar donors.
CC It is specific for D-glycerate, but it can also use D-lactate and
CC glycolate as sugar acceptors. This reaction occurs with a net retention
CC of anomeric configuration; the newly formed glycosidic linkage has the
CC same alpha configuration as the sugar donor.
CC {ECO:0000269|PubMed:10585410, ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(R)-glycerate + GDP-alpha-D-mannose = (2R)-2-O-(alpha-D-
CC mannosyl)-glycerate + GDP + H(+); Xref=Rhea:RHEA:30639,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:16659, ChEBI:CHEBI:57527,
CC ChEBI:CHEBI:57541, ChEBI:CHEBI:58189; EC=2.4.1.269;
CC Evidence={ECO:0000269|PubMed:10585410, ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:21288903};
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:21288903};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:21288903};
CC Name=Ni(2+); Xref=ChEBI:CHEBI:49786;
CC Evidence={ECO:0000269|PubMed:21288903};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:21288903};
CC Note=Divalent cations such as magnesium, calcium and to a lesser extent
CC manganese, nickel and cobalt. {ECO:0000269|PubMed:21288903};
CC -!- ACTIVITY REGULATION: Inhibited by GDP. {ECO:0000269|PubMed:21288903}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.2 uM for D-glycerate {ECO:0000269|PubMed:10585410,
CC ECO:0000269|PubMed:15951819, ECO:0000269|PubMed:21288903};
CC KM=3.1 uM for GDP-Man {ECO:0000269|PubMed:10585410,
CC ECO:0000269|PubMed:15951819, ECO:0000269|PubMed:21288903};
CC KM=81.2 uM for GDP-Man (in the presence of calcium ions at 25 degrees
CC Celsius) {ECO:0000269|PubMed:10585410, ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903};
CC KM=89.4 uM for GDP-Man (in the presence of calcium ions at 65 degrees
CC Celsius) {ECO:0000269|PubMed:10585410, ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903};
CC KM=96.5 uM for D-glycerate (in the presence of calcium ions at 65
CC degrees Celsius) {ECO:0000269|PubMed:10585410,
CC ECO:0000269|PubMed:15951819, ECO:0000269|PubMed:21288903};
CC KM=121.9 uM for D-glycerate (in the presence of calcium ions at 25
CC degrees Celsius) {ECO:0000269|PubMed:10585410,
CC ECO:0000269|PubMed:15951819, ECO:0000269|PubMed:21288903};
CC KM=124.9 uM for GDP-Glc (in the presence of calcium ions at 25
CC degrees Celsius) {ECO:0000269|PubMed:10585410,
CC ECO:0000269|PubMed:15951819, ECO:0000269|PubMed:21288903};
CC KM=138.6 uM for GDP-Glc (in the presence of calcium ions at 65
CC degrees Celsius) {ECO:0000269|PubMed:10585410,
CC ECO:0000269|PubMed:15951819, ECO:0000269|PubMed:21288903};
CC KM=300 uM for GDP-Man (at 90 degrees Celsius)
CC {ECO:0000269|PubMed:10585410, ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903};
CC KM=600 uM for D-glycerate (at 90 degrees Celsius (PubMed 10585410))
CC {ECO:0000269|PubMed:10585410, ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903};
CC Note=kcat is 6.5 sec(-1) for mannosylglycerate synthase activity with
CC D-glycerate as sugar acceptor (in the presence of calcium ions at 65
CC degrees Celsius) (PubMed:15951819). kcat is 6.1 sec(-1) for
CC mannosylglycerate synthase activity with GDP-Man as sugar donor (in
CC the presence of calcium ions at 65 degrees Celsius)
CC (PubMed:15951819). kcat is 1.9 sec(-1) for mannosylglycerate synthase
CC activity with D-glycerate (PubMed:21288903). kcat is 1.1 sec(-1) for
CC mannosylglycerate synthase activity with GDP-Glc as sugar donor (in
CC the presence of calcium ions at 65 degrees Celsius)
CC (PubMed:15951819). kcat is 1.02 sec(-1) for mannosylglycerate
CC synthase activity with GDP-Man and D-glycerate as sugar donor and
CC acceptor, respectively (in the presence of calcium ions at 25 degrees
CC Celsius) (PubMed:15951819). kcat is 0.63 sec(-1) for
CC mannosylglycerate synthase activity with GDP-Glc as sugar donor (in
CC the presence of calcium ions at 25 degrees Celsius)
CC (PubMed:15951819). {ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903};
CC pH dependence:
CC Optimum pH is between 5.5 and 7. {ECO:0000269|PubMed:10585410,
CC ECO:0000269|PubMed:15951819, ECO:0000269|PubMed:21288903};
CC Temperature dependence:
CC Optimum temperature is between 85 and 90 degrees Celsius. It is
CC stable at high temperatures, however at 90 degrees Celsius the
CC stability of the enzyme is only moderate and 50% of the activity is
CC lost after 30 minutes incubation. At 65 degrees Celsius, the specific
CC enzymatic activity is 5-fold lower than the maximum value.
CC {ECO:0000269|PubMed:10585410, ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903};
CC -!- SUBUNIT: Homotetramer. Dimer of dimers. {ECO:0000269|PubMed:15951819,
CC ECO:0000269|PubMed:21288903}.
CC -!- INTERACTION:
CC Q9RFR0; Q9RFR0: mgs; NbExp=3; IntAct=EBI-15553684, EBI-15553684;
CC -!- MISCELLANEOUS: The biosynthesis of mannosylglycerate in R.marinus can
CC proceed via an other alternative pathway, in wich a mannosyl-3-
CC phosphoglycerate synthase catalyzes the conversion of GDP mannose and
CC D-3-phosphoglycerate into a phosphorylated intermediate, which is
CC subsequently converted to mannosylglycerate by the action of a
CC mannosyl-3-phosphoglycerate phosphatase. A noticeable feature
CC distinguishing the two enzymatic systems involved in the synthesis of
CC MG is their dependence on salt; while the mannosylglycerate synthase
CC reaction is salt-independent, addition of NaCl or KCl is required to
CC achieve full activity of the mannosyl-3-phosphoglycerate
CC synthase/phosphatase system (PubMed:10585410).
CC {ECO:0000305|PubMed:10585410}.
CC -!- SIMILARITY: Belongs to the glycosyltransferase 78 family.
CC {ECO:0000305}.
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DR EMBL; AF173987; AAF16905.1; -; Genomic_DNA.
DR PDB; 2BO4; X-ray; 1.95 A; A/B/C/D/E/F=1-397.
DR PDB; 2BO6; X-ray; 2.45 A; A/B=1-397.
DR PDB; 2BO7; X-ray; 2.95 A; A/B/C/D/E/F/G/H/I/J=1-397.
DR PDB; 2BO8; X-ray; 2.80 A; A/B/C/D/E/F/G/H/I/J=1-397.
DR PDB; 2Y4J; X-ray; 2.30 A; A/B=1-382.
DR PDBsum; 2BO4; -.
DR PDBsum; 2BO6; -.
DR PDBsum; 2BO7; -.
DR PDBsum; 2BO8; -.
DR PDBsum; 2Y4J; -.
DR AlphaFoldDB; Q9RFR0; -.
DR SMR; Q9RFR0; -.
DR DIP; DIP-60494N; -.
DR CAZy; GT78; Glycosyltransferase Family 78.
DR BioCyc; MetaCyc:MON-13377; -.
DR BRENDA; 2.4.1.269; 5425.
DR SABIO-RK; Q9RFR0; -.
DR EvolutionaryTrace; Q9RFR0; -.
DR PRO; PR:Q9RFR0; -.
DR GO; GO:0016758; F:hexosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0042802; F:identical protein binding; IPI:IntAct.
DR GO; GO:0102921; F:mannosylglycerate synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IDA:UniProtKB.
DR GO; GO:0051479; P:mannosylglycerate biosynthetic process; IDA:CACAO.
DR Gene3D; 3.90.550.10; -; 2.
DR InterPro; IPR029044; Nucleotide-diphossugar_trans.
DR SUPFAM; SSF53448; SSF53448; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Cobalt; Direct protein sequencing; Magnesium;
KW Manganese; Metal-binding; Nickel; Transferase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:10585410"
FT CHAIN 2..397
FT /note="Mannosylglycerate synthase"
FT /id="PRO_0000429861"
FT BINDING 7..11
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 35
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 66
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 76
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 100..101
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 100
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 102
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT BINDING 131
FT /ligand="(R)-glycerate"
FT /ligand_id="ChEBI:CHEBI:16659"
FT BINDING 136..139
FT /ligand="(R)-glycerate"
FT /ligand_id="ChEBI:CHEBI:16659"
FT BINDING 163
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 192
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 217
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT BINDING 218
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT BINDING 220
FT /ligand="GDP-alpha-D-mannose"
FT /ligand_id="ChEBI:CHEBI:57527"
FT MUTAGEN 9
FT /note="K->A: The catalytic efficiency is almost one order
FT of magnitude higher than wild-type enzyme for both
FT substrates. The mutation has little effect on the affinity
FT binding value for GDP-Man and D-glycerate."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 11
FT /note="E->A: Results in a modest increase in the catalytic
FT efficiency coupled with a decrease in the affinity binding
FT value for GDP-Man."
FT /evidence="ECO:0000269|PubMed:15951819"
FT MUTAGEN 37
FT /note="Y->A: Significant increase in catalytic efficiency.
FT The mutation has little effect on the affinity binding
FT value for GDP-Man, however it shows an 4-fold decrease in
FT the affinity binding value for D-glycerate."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 66
FT /note="Q->A: Results in an increase in the catalytic
FT efficiency (up to 72-fold) coupled with a decrease in the
FT affinity binding for both D-glycerate and GDP-Man."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 76
FT /note="K->A: Results in a 3-fold increase in the catalytic
FT efficiency coupled with a decrease in affinity binding for
FT D-glycerate and GDP-Man of 15- and 3-fold, respectively."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 100
FT /note="D->A: Completely inactive."
FT /evidence="ECO:0000269|PubMed:15951819"
FT MUTAGEN 102
FT /note="D->A: Completely inactive."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 131
FT /note="R->A: Completely inactive."
FT /evidence="ECO:0000269|PubMed:15951819"
FT MUTAGEN 135
FT /note="D->A: Results in a extremely low affinity binding
FT value for D-glycerate (5600-fold lower than wild-type) and
FT displays a slight increase in the catalytic efficiency (2-
FT fold) compared with wild-type enzyme."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 139
FT /note="T->A: Results in a modest decrease in the catalytic
FT efficiency coupled with a 1500-fold decrease in the
FT affinity binding value for D-glycerate. The mutant is 500-
FT fold less active when D-lactate, rather than D-glycerate,
FT is the acceptor substrate."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 189
FT /note="W->A: Results in a 3-fold increase in the catalytic
FT efficiency coupled with a 7-fold decrease in the affinity
FT binding for D-glycerate compared with the wild-type enzyme.
FT It does not influence utilization of GDP-Man."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 192
FT /note="D->A: Completely inactive."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 217
FT /note="H->A: Displays a significant change in metal
FT preference. This mutant is essentially inactive in the
FT presence of calcium ions (specific activity 1000-fold lower
FT than wild-type), whereas the catalytic efficiency value is
FT 23-fold lower in the presence of magnesium ions."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 218
FT /note="R->A: Results in a significant increase in the
FT catalytic efficiency coupled with a decrease in the
FT affinity binding value for GDP-Man. It has only a modest
FT influence on the affinity binding value for D-glycerate."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 220
FT /note="Y->A: Results in a 500-fold decrease in the affinity
FT binding value for D-glycerate in the presence of saturating
FT GDP-Man."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 220
FT /note="Y->F: Results in a 1500-fold decrease in the
FT affinity binding value for D-glycerate in the presence of
FT saturating GDP-Man."
FT /evidence="ECO:0000269|PubMed:21288903"
FT MUTAGEN 229
FT /note="M->A: Minor influence."
FT /evidence="ECO:0000269|PubMed:21288903"
FT STRAND 3..7
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 13..25
FT /evidence="ECO:0007829|PDB:2BO4"
FT STRAND 31..38
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 41..57
FT /evidence="ECO:0007829|PDB:2BO4"
FT STRAND 61..65
FT /evidence="ECO:0007829|PDB:2BO4"
FT STRAND 70..75
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 76..90
FT /evidence="ECO:0007829|PDB:2BO4"
FT STRAND 94..98
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 108..119
FT /evidence="ECO:0007829|PDB:2BO4"
FT STRAND 123..128
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 137..141
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 143..150
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 156..158
FT /evidence="ECO:0007829|PDB:2BO4"
FT STRAND 167..170
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 171..179
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 181..184
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 191..201
FT /evidence="ECO:0007829|PDB:2BO4"
FT STRAND 206..210
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 223..226
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 227..240
FT /evidence="ECO:0007829|PDB:2BO4"
FT STRAND 251..253
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 261..264
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 271..278
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 284..289
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 290..292
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 295..303
FT /evidence="ECO:0007829|PDB:2BO4"
FT TURN 304..306
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 315..328
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 334..354
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 356..358
FT /evidence="ECO:0007829|PDB:2BO4"
FT HELIX 360..380
FT /evidence="ECO:0007829|PDB:2BO4"
SQ SEQUENCE 397 AA; 46126 MW; 10303BFBF298E6EC CRC64;
MSLVVFPFKH EHPEVLLHNV RVAAAHPRVH EVLCIGYERD QTYEAVERAA PEISRATGTP
VSVRLQERLG TLRPGKGDGM NTALRYFLEE TQWERIHFYD ADITSFGPDW ITKAEEAADF
GYGLVRHYFP RASTDAMITW MITRTGFALL WPHTELSWIE QPLGGELLMR REVAAMLYED
ERVRRRSDWG IDTLYTFVTV QQGVSIYECY IPEGKAHRLY GGLDDLRTML VECFAAIQSL
QHEVVGQPAI HRQEHPHRVP VHIAERVGYD VEATLHRLMQ HWTPRQVELL ELFTTPVREG
LRTCQRRPAF NFMDEMAWAA TYHVLLEHFQ PGDPDWEELL FKLWTTRVLN YTMTVALRGY
DYAQQYLYRM LGRYRYQAAL ENGRGHPVPP RAALSTA
