BRENDA - Enzyme Database show
show all sequences of 2.4.1.65

Purification, kinetic characterization, and mapping of the minimal catalytic domain and the key polar goups of Helicobacter pylori alpha-(1,3/1,4)-fucosyltransferases

Ma, B.; Audette, G.F.; Lin, S.; Palcic, M.M.; Hazes, B.; Taylor, D.E.; J. Biol. Chem. 281, 6385-6394 (2006)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
full-length and each truncated FucT (DELTAFucT) are expressed in Escherichia coli HMS174DE3
Helicobacter pylori
Engineering
Amino acid exchange
Commentary
Organism
additional information
with full length UA948FucT, only 20% of total enzyme activities are localized in the soluble fraction. For UA948(1-428), this increases to 47%, respectively, indicating that truncation of the C-terminal putative alpha-helices increases FucT solubility. UA948(1-364), a mutant with the entire heptad repeat region removed, exhibits extremely low levels of enzyme activity. This indicates that the heptad repeat region is essential for enzyme activity. Construct UA948(1-371) has little enzyme activity. UA948(10-434) and UA948(1-434) completely lose activity
Helicobacter pylori
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0119
-
GDP-beta-L-fucose
cosubstrate: Galbeta1,3GlcNAc-O(CH2)8CO2CH3, truncated enzyme UA948(1-428)
Helicobacter pylori
0.0337
-
GDP-beta-L-fucose
cosubstrate: Galbeta1,3GlcNAc-O(CH2)8CO2CH3, full-length enzyme
Helicobacter pylori
5.7
-
Galbeta1,3GlcNAc-O(CH2)8CO2CH3
truncated enzyme UA948(1-428)
Helicobacter pylori
13.3
-
Galbeta1,3GlcNAc-O(CH2)8CO2CH3
full-length enzyme
Helicobacter pylori
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
51200
-
51200, UA948FucT(1-428), calculated from sequence
Helicobacter pylori
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Helicobacter pylori
-
UA948
-
Helicobacter pylori UA948
-
UA948
-
Purification (Commentary)
Commentary
Organism
UA948FucT(1-428) with a His6 tag at the C terminus is purified
Helicobacter pylori
Storage Stability
Storage Stability
Organism
-80°C, 3 months, lyophilized UA948FucT(1-428) loses 23% of its activity
Helicobacter pylori
most stable at -80°C with or without glycerol supplementation
Helicobacter pylori
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
GDP-beta-L-fucose + 3-sulfo-Galbeta1,3GlcNAc
40% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori
?
-
-
-
?
GDP-beta-L-fucose + 3-sulfo-Galbeta1,3GlcNAc
40% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori NCTC11639
?
-
-
-
?
GDP-beta-L-fucose + 3-sulfo-Galbeta1,3GlcNAc
40% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori UA948
?
-
-
-
?
GDP-beta-L-fucose + Fucalpha1,2Galbeta1,3GlcNAc
34% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori
?
-
-
-
?
GDP-beta-L-fucose + Fucalpha1,2Galbeta1,3GlcNAc
34% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori NCTC11639
?
-
-
-
?
GDP-beta-L-fucose + Fucalpha1,2Galbeta1,3GlcNAc
34% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori UA948
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc
-
680736
Helicobacter pylori
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc
-
680736
Helicobacter pylori NCTC11639
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc
-
680736
Helicobacter pylori UA948
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc-O(CH2)8CO2CH3
-
680736
Helicobacter pylori
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc-O(CH2)8CO2CH3
-
680736
Helicobacter pylori NCTC11639
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc-O(CH2)8CO2CH3
-
680736
Helicobacter pylori UA948
?
-
-
-
?
additional information
the enzyme shows alpha1,3- and alpha1,4-fucosyltransferase activity
680736
Helicobacter pylori
?
-
-
-
-
additional information
the enzyme shows alpha1,3- and alpha1,4-fucosyltransferase activity
680736
Helicobacter pylori NCTC11639
?
-
-
-
-
additional information
the enzyme shows alpha1,3- and alpha1,4-fucosyltransferase activity
680736
Helicobacter pylori UA948
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
?
51200, UA948FucT(1-428), calculated from sequence
Helicobacter pylori
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1
-
Galbeta1,3GlcNAc-O(CH2)8CO2CH3
truncated enzyme UA948(1-428)
Helicobacter pylori
1.1
-
GDP-beta-L-fucose
cosubstrate: Galbeta1,3GlcNAc-O(CH2)8CO2CH3, truncated enzyme UA948(1-428)
Helicobacter pylori
Cloned(Commentary) (protein specific)
Commentary
Organism
full-length and each truncated FucT (DELTAFucT) are expressed in Escherichia coli HMS174DE3
Helicobacter pylori
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
with full length UA948FucT, only 20% of total enzyme activities are localized in the soluble fraction. For UA948(1-428), this increases to 47%, respectively, indicating that truncation of the C-terminal putative alpha-helices increases FucT solubility. UA948(1-364), a mutant with the entire heptad repeat region removed, exhibits extremely low levels of enzyme activity. This indicates that the heptad repeat region is essential for enzyme activity. Construct UA948(1-371) has little enzyme activity. UA948(10-434) and UA948(1-434) completely lose activity
Helicobacter pylori
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0119
-
GDP-beta-L-fucose
cosubstrate: Galbeta1,3GlcNAc-O(CH2)8CO2CH3, truncated enzyme UA948(1-428)
Helicobacter pylori
0.0337
-
GDP-beta-L-fucose
cosubstrate: Galbeta1,3GlcNAc-O(CH2)8CO2CH3, full-length enzyme
Helicobacter pylori
5.7
-
Galbeta1,3GlcNAc-O(CH2)8CO2CH3
truncated enzyme UA948(1-428)
Helicobacter pylori
13.3
-
Galbeta1,3GlcNAc-O(CH2)8CO2CH3
full-length enzyme
Helicobacter pylori
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
51200
-
51200, UA948FucT(1-428), calculated from sequence
Helicobacter pylori
Purification (Commentary) (protein specific)
Commentary
Organism
UA948FucT(1-428) with a His6 tag at the C terminus is purified
Helicobacter pylori
Storage Stability (protein specific)
Storage Stability
Organism
-80°C, 3 months, lyophilized UA948FucT(1-428) loses 23% of its activity
Helicobacter pylori
most stable at -80°C with or without glycerol supplementation
Helicobacter pylori
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
GDP-beta-L-fucose + 3-sulfo-Galbeta1,3GlcNAc
40% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori
?
-
-
-
?
GDP-beta-L-fucose + 3-sulfo-Galbeta1,3GlcNAc
40% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori NCTC11639
?
-
-
-
?
GDP-beta-L-fucose + 3-sulfo-Galbeta1,3GlcNAc
40% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori UA948
?
-
-
-
?
GDP-beta-L-fucose + Fucalpha1,2Galbeta1,3GlcNAc
34% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori
?
-
-
-
?
GDP-beta-L-fucose + Fucalpha1,2Galbeta1,3GlcNAc
34% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori NCTC11639
?
-
-
-
?
GDP-beta-L-fucose + Fucalpha1,2Galbeta1,3GlcNAc
34% of the activity with Galbeta1,3GlcNAc
680736
Helicobacter pylori UA948
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc
-
680736
Helicobacter pylori
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc
-
680736
Helicobacter pylori NCTC11639
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc
-
680736
Helicobacter pylori UA948
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc-O(CH2)8CO2CH3
-
680736
Helicobacter pylori
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc-O(CH2)8CO2CH3
-
680736
Helicobacter pylori NCTC11639
?
-
-
-
?
GDP-beta-L-fucose + Galbeta1,3GlcNAc-O(CH2)8CO2CH3
-
680736
Helicobacter pylori UA948
?
-
-
-
?
additional information
the enzyme shows alpha1,3- and alpha1,4-fucosyltransferase activity
680736
Helicobacter pylori
?
-
-
-
-
additional information
the enzyme shows alpha1,3- and alpha1,4-fucosyltransferase activity
680736
Helicobacter pylori NCTC11639
?
-
-
-
-
additional information
the enzyme shows alpha1,3- and alpha1,4-fucosyltransferase activity
680736
Helicobacter pylori UA948
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
?
51200, UA948FucT(1-428), calculated from sequence
Helicobacter pylori
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1
-
Galbeta1,3GlcNAc-O(CH2)8CO2CH3
truncated enzyme UA948(1-428)
Helicobacter pylori
1.1
-
GDP-beta-L-fucose
cosubstrate: Galbeta1,3GlcNAc-O(CH2)8CO2CH3, truncated enzyme UA948(1-428)
Helicobacter pylori
Other publictions for EC 2.4.1.65
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
736817
Huang
Sperm fucosyltransferase-5 med ...
Homo sapiens
Mol. Hum. Reprod.
21
516-526
2015
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1
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1
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2
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1
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4
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1
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1
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1
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1
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4
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1
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2
2
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735774
Seelhorst
Synthesis and analysis of pote ...
Helicobacter pylori, Homo sapiens
Bioorg. Med. Chem.
22
6430-6437
2014
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22
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2
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7
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7
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7
22
7
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735422
Wang
alpha(1,3)-Fucosyltransferases ...
Mus musculus
Am. J. Pathol.
182
2082-2093
2013
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-
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1
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1
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1
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-
-
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-
3
3
-
-
-
735957
Lunau
Fluorescently labeled substrat ...
Helicobacter pylori, Homo sapiens
Chemistry
19
17379-17390
2013
-
-
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1
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2
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2
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1
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2
-
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-
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736194
Wan
Fucosyltransferase VII improve ...
Mus musculus
Glycobiology
23
1184-1191
2013
-
-
-
-
-
-
-
-
-
-
-
1
-
3
-
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-
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4
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2
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1
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4
-
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2
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-
-
-
-
-
-
-
-
1
1
-
-
-
736582
Brazil
alpha3/4 Fucosyltransferase 3- ...
Homo sapiens
J. Immunol.
191
4804-4817
2013
-
-
-
-
-
-
-
-
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-
1
-
4
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5
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1
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1
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5
-
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1
-
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-
-
-
-
-
-
-
-
1
1
-
-
-
736601
Saeki
alpha(1,3) fucosyltransferases ...
Mus musculus
J. Invest. Dermatol.
133
2161-2169
2013
-
-
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-
-
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2
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1
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1
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-
1
1
-
-
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721501
Guo
Functional analysis of alpha1, ...
Homo sapiens
Biochem. Biophys. Res. Commun.
417
311-317
2012
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1
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3
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3
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-
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1
1
1
1
-
-
704049
Carvalho
Differential expression of alp ...
Homo sapiens
Int. J. Biochem. Cell Biol.
42
80-89
2010
-
-
-
-
-
-
-
-
-
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1
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8
-
-
14
-
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8
-
-
14
-
-
-
-
-
-
-
-
-
2
1
1
2
-
-
703056
Tefsen
Chemoenzymatic synthesis of mu ...
Homo sapiens
Carbohydr. Res.
344
1501-1507
2009
-
-
1
-
-
-
-
4
-
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4
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1
4
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1
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4
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1
4
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1
1
-
-
-
703893
Fagerberg
Novel Leb-like Helicobacter py ...
Homo sapiens
Glycobiology
19
182-191
2009
-
-
1
-
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-
-
-
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6
-
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3
-
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1
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3
-
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-
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-
1
1
-
-
-
704613
Mollicone
Activity, splice variants, con ...
Homo sapiens
J. Biol. Chem.
284
4723-4738
2009
-
-
1
-
-
-
-
4
3
1
-
-
-
2
-
-
-
-
-
17
-
-
4
-
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2
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4
4
2
-
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17
-
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4
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
706522
Barthel
Alpha 1,3 fucosyltransferases ...
Homo sapiens
Proc. Natl. Acad. Sci. USA
106
19491-19496
2009
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1
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2
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2
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-
-
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-
2
1
1
2
-
-
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Molecular cloning and function ...
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1
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Sousa
Deletion of the cytoplasmic do ...
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2004
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661313
Palma
Effect of the manganese ion on ...
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2004
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661341
Norris
Inhibition kinetics of carba- ...
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2004
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3
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3
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1
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Abe
Yeast cells harboring human al ...
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Galan
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Stoykova
alpha1,3 Fucosyltransferases i ...
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Chandrasekaran
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alpha1,4-Fucosyltransferase ac ...
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