complex with tripolyphosphate, to 2.3 A resolution and docking solution of substrate thiamine triphosphate. The thiazole ring of the thiamine forms a stacking interaction with Trp53, and the aminopyrimidine ring lies in a pocket defined by His76, Pro191 and Ile195. The triphosphate moiety occupies a similar position as in the enzyme-tripolyphosphate complex, with interactions involving the side chains of Lys11, Arg55, Arg57, Lys65, Arg125 and Lys193
Homo sapiens
D147A
about 30% of wild-type catalytic efficiency
Mus musculus
D37A
about 0.5% of wild-type catalytic efficiency
Mus musculus
E81A
about 30% of wild-type catalytic efficiency
Mus musculus
K11A
about 15% of wild-type catalytic efficiency
Mus musculus
K193A
about 60% of wild-type catalytic efficiency
Mus musculus
K65A
more than 1000fold decrease in kcat value
Mus musculus
W53A
2.5fold increase in catalytic efficiency
Mus musculus
Y39A
about 30% of wild-type catalytic efficiency
Mus musculus
Y79A
about 5% of wild-type catalytic efficiency
Mus musculus
0.008
thiamine triphosphate
wild-type, pH 8.5, 37°C
Mus musculus
0.009
thiamine triphosphate
mutant K65A, pH 8.5, 37°C
Mus musculus
0.012
thiamine triphosphate
mutant K11A, pH 8.5, 37°C
Mus musculus
0.013
thiamine triphosphate
mutant E81A, pH 8.5, 37°C
Mus musculus
0.042
thiamine triphosphate
mutant Y79A, pH 8.5, 37°C
Mus musculus
0.2
thiamine triphosphate
mutant D37A, pH 8.5, 37°C
Mus musculus
0.55
thiamine triphosphate
mutant W53A, pH 8.5, 37°C
Mus musculus
0.62
thiamine triphosphate
mutant Y39A, pH 8.5, 37°C
Mus musculus
0.74
thiamine triphosphate
mutant K193A, pH 8.5, 37°C
Mus musculus
2.8
thiamine triphosphate
mutant D147A, pH 8.5, 37°C
Mus musculus
0.019
thiamine triphosphate
mutant K65A, pH 8.5, 37°C
Mus musculus
0.64
thiamine triphosphate
mutant Y39A, pH 8.5, 37°C
Mus musculus
1.5
thiamine triphosphate
mutant K193A, pH 8.5, 37°C
Mus musculus
2.6
thiamine triphosphate
mutant D147A, pH 8.5, 37°C
Mus musculus
2.8
thiamine triphosphate
mutant D37A, pH 8.5, 37°C
Mus musculus
4.4
thiamine triphosphate
mutant W53A, pH 8.5, 37°C
Mus musculus
6
thiamine triphosphate
mutant K11A, pH 8.5, 37°C; mutant Y79A, pH 8.5, 37°C
Mus musculus
14.1
thiamine triphosphate
mutant E81A, pH 8.5, 37°C
Mus musculus
23
thiamine triphosphate
wild-type, pH 8.5, 37°C
Mus musculus
complex with tripolyphosphate, to 2.3 A resolution and docking solution of substrate thiamine triphosphate. The thiazole ring of the thiamine forms a stacking interaction with Trp53, and the aminopyrimidine ring lies in a pocket defined by His76, Pro191 and Ile195. The triphosphate moiety occupies a similar position as in the enzyme-tripolyphosphate complex, with interactions involving the side chains of Lys11, Arg55, Arg57, Lys65, Arg125 and Lys193
Homo sapiens
D147A
about 30% of wild-type catalytic efficiency
Mus musculus
D37A
about 0.5% of wild-type catalytic efficiency
Mus musculus
E81A
about 30% of wild-type catalytic efficiency
Mus musculus
K11A
about 15% of wild-type catalytic efficiency
Mus musculus
K193A
about 60% of wild-type catalytic efficiency
Mus musculus
K65A
more than 1000fold decrease in kcat value
Mus musculus
W53A
2.5fold increase in catalytic efficiency
Mus musculus
Y39A
about 30% of wild-type catalytic efficiency
Mus musculus
Y79A
about 5% of wild-type catalytic efficiency
Mus musculus
0.008
thiamine triphosphate
wild-type, pH 8.5, 37°C
Mus musculus
0.009
thiamine triphosphate
mutant K65A, pH 8.5, 37°C
Mus musculus
0.012
thiamine triphosphate
mutant K11A, pH 8.5, 37°C
Mus musculus
0.013
thiamine triphosphate
mutant E81A, pH 8.5, 37°C
Mus musculus
0.042
thiamine triphosphate
mutant Y79A, pH 8.5, 37°C
Mus musculus
0.2
thiamine triphosphate
mutant D37A, pH 8.5, 37°C
Mus musculus
0.55
thiamine triphosphate
mutant W53A, pH 8.5, 37°C
Mus musculus
0.62
thiamine triphosphate
mutant Y39A, pH 8.5, 37°C
Mus musculus
0.74
thiamine triphosphate
mutant K193A, pH 8.5, 37°C
Mus musculus
2.8
thiamine triphosphate
mutant D147A, pH 8.5, 37°C
Mus musculus
0.019
thiamine triphosphate
mutant K65A, pH 8.5, 37°C
Mus musculus
0.64
thiamine triphosphate
mutant Y39A, pH 8.5, 37°C
Mus musculus
1.5
thiamine triphosphate
mutant K193A, pH 8.5, 37°C
Mus musculus
2.6
thiamine triphosphate
mutant D147A, pH 8.5, 37°C
Mus musculus
2.8
thiamine triphosphate
mutant D37A, pH 8.5, 37°C
Mus musculus
4.4
thiamine triphosphate
mutant W53A, pH 8.5, 37°C
Mus musculus
6
thiamine triphosphate
mutant K11A, pH 8.5, 37°C; mutant Y79A, pH 8.5, 37°C
Mus musculus
14.1
thiamine triphosphate
mutant E81A, pH 8.5, 37°C
Mus musculus
23
thiamine triphosphate
wild-type, pH 8.5, 37°C
Mus musculus
14
thiamine triphosphate
mutant D37A, pH 8.5, 37°C
Mus musculus
140
thiamine triphosphate
mutant Y79A, pH 8.5, 37°C
Mus musculus
500
thiamine triphosphate
mutant K11A, pH 8.5, 37°C
Mus musculus
1000
thiamine triphosphate
mutant D147A, pH 8.5, 37°C; mutant E81A, pH 8.5, 37°C; mutant Y39A, pH 8.5, 37°C
Mus musculus
2000
thiamine triphosphate
mutant K193A, pH 8.5, 37°C; mutant K65A, pH 8.5, 37°C
Mus musculus
3000
thiamine triphosphate
wild-type, pH 8.5, 37°C
Mus musculus
8000
thiamine triphosphate
mutant W53A, pH 8.5, 37°C
Mus musculus
14
thiamine triphosphate
mutant D37A, pH 8.5, 37°C
Mus musculus
140
thiamine triphosphate
mutant Y79A, pH 8.5, 37°C
Mus musculus
500
thiamine triphosphate
mutant K11A, pH 8.5, 37°C
Mus musculus
1000
thiamine triphosphate
mutant D147A, pH 8.5, 37°C; mutant E81A, pH 8.5, 37°C; mutant Y39A, pH 8.5, 37°C
Mus musculus
2000
thiamine triphosphate
mutant K193A, pH 8.5, 37°C; mutant K65A, pH 8.5, 37°C
Mus musculus
3000
thiamine triphosphate
wild-type, pH 8.5, 37°C
Mus musculus
8000
thiamine triphosphate
mutant W53A, pH 8.5, 37°C
Mus musculus
751047
Martinez
Structural determinants for s ...
Mus musculus
J. Biol. Chem.
290
23348-23360
2015
-
-
-
1
-
-
-
-
-
1
-
1
-
3
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
733393
Delvaux
Structural determinants of spe ...
Homo sapiens, Mus musculus
Biochim. Biophys. Acta
1830
4513-4523
2013
-
-
2
1
9
-
-
10
-
1
-
-
-
2
-
-
-
-
-
-
-
-
1
-
1
-
-
-
9
-
-
-
-
-
-
-
-
-
2
-
1
9
-
-
-
-
10
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
9
-
-
-
-
-
-
-
-
7
7
687693
Song
Structural basis for the catal ...
Mus musculus
J. Biol. Chem.
283
10939-10948
2008
-
-
-
-
-
-
1
1
1
1
1
1
-
2
-
-
-
1
-
-
-
-
2
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
1
1
1
1
-
-
-
-
-
-
-
-
2
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
696404
Kovacevic
The iron-regulated metastasis ...
Homo sapiens, Rattus norvegicus
Biochim. Biophys. Acta
1783
1981-1992
2008
2
-
2
-
2
-
-
-
-
-
-
2
-
2
-
-
-
-
-
3
-
-
4
-
4
-
-
-
-
-
-
-
-
-
-
-
2
-
2
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
3
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
667797
Szyniarowski Piot
Pig tissues express a catalyti ...
Homo sapiens, Sus scrofa
Biochim. Biophys. Acta
1725
93-102
2005
-
-
2
-
4
-
2
3
-
5
2
-
-
6
-
-
2
-
-
1
16
-
2
-
4
-
-
1
-
5
-
-
-
2
-
1
-
-
2
-
-
4
-
1
2
2
3
-
5
2
-
-
-
-
2
-
1
16
-
2
-
-
-
1
-
5
-
-
-
-
-
-
-
-
-
655738
Lakaye
Human recombinant thiamine tri ...
Homo sapiens
Int. J. Biochem. Cell Biol.
36
1348-1364
2004
-
-
1
-
-
-
6
2
1
2
-
-
-
3
-
-
1
-
-
-
1
-
2
-
-
-
-
2
2
1
1
-
-
2
-
1
-
-
1
-
-
-
-
1
7
2
2
1
2
-
-
-
-
-
1
-
-
1
-
2
-
-
-
2
2
1
1
-
-
-
-
-
-
-
-
656873
Czerniecki
Neuronal localization of the 2 ...
Rattus norvegicus
Neuroscience
125
833-840
2004
-
-
-
-
-
-
-
-
1
-
-
1
-
4
-
-
-
-
-
3
-
-
2
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
3
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
668963
Lakaye
Expression of 25 kDa thiamine ...
Mus musculus, Rattus norvegicus
Int. J. Biochem. Cell Biol.
36
2032-2041
2004
-
-
-
-
-
-
-
1
-
-
-
-
-
5
-
-
-
-
-
16
-
-
2
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
16
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
655198
Makarchikov
Thiamine triphosphate and thia ...
Bos taurus, Gallus gallus, Homo sapiens, quail, Rattus norvegicus, Sus scrofa
Cell. Mol. Life Sci.
60
1477-1488
2003
-
-
-
-
-
-
-
15
-
-
17
-
-
12
-
-
-
-
-
25
22
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
15
-
-
17
-
-
-
-
-
-
25
22
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
656042
Lakaye
Molecular characterization of ...
Bos taurus, Homo sapiens
J. Biol. Chem.
277
13771-13777
2002
-
-
1
-
-
-
4
2
1
-
6
-
-
7
-
-
2
-
-
9
2
-
4
2
2
-
-
-
2
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
4
-
2
1
-
6
-
-
-
-
2
-
9
2
-
4
2
-
-
-
2
-
-
-
-
-
-
-
-
-
-
210023
Makarchikov
Thiamine triphosphatase activi ...
Bos taurus
Biochem. Mol. Biol. Int.
46
115-123
1998
-
-
-
-
-
-
-
2
-
-
1
-
-
2
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
1
-
-
2
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
210021
Bettendorff
Thiamine, thiamine phosphates, ...
Homo sapiens
J. Neurochem.
66
250-258
1996
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
210020
Makarchikov
Purification and characterizat ...
Bos taurus
Biochim. Biophys. Acta
1117
326-332
1992
-
-
-
-
-
1
-
1
1
6
2
-
-
2
-
-
1
-
-
2
1
2
7
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
1
6
2
-
-
-
-
1
-
2
1
2
7
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
210012
Matsuda
Membrane-associated thiamine t ...
Rattus norvegicus
Int. J. Biochem.
23
1111-1114
1991
-
-
-
-
-
-
9
-
1
-
-
-
-
2
-
-
-
-
-
2
-
-
3
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
9
-
-
1
-
-
-
-
-
-
-
-
2
-
-
3
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
210022
Bettendorff
Solubilization and thiamine tr ...
Electrophorus electricus
Biochim. Biophys. Acta
1073
69-76
1991
4
-
-
-
-
-
3
-
1
-
-
-
-
2
-
-
-
-
-
2
-
1
1
-
-
-
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
3
-
-
1
-
-
-
-
-
-
-
-
2
-
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
210024
Bettendorff
Application of high-performanc ...
Electrophorus electricus
J. Chromatogr.
566
397-408
1991
-
-
-
-
-
-
-
-
-
1
-
1
-
3
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210019
Nishimune
Hydrolysis and synthesis of th ...
Aneurinibacillus aneurinilyticus, Escherichia coli, Escherichia coli W 1485
J. Nutr. Sci. Vitaminol.
33
113-127
1987
-
-
-
-
-
-
3
2
2
1
-
-
-
3
-
-
-
-
-
-
-
-
3
-
-
2
-
-
-
2
-
-
-
1
-
-
-
-
-
-
-
-
-
-
3
1
2
2
1
-
-
-
-
-
-
-
-
-
-
3
-
2
-
-
-
2
-
-
-
-
-
-
-
-
-
210018
Ogawa
Recent findings on ultracytoch ...
Rattus norvegicus
Ann. N. Y. Acad. Sci.
378
188-214
1982
-
-
-
-
-
-
-
-
4
-
-
1
-
1
-
-
-
-
-
3
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
1
-
-
-
-
-
3
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210017
Penttinen
The relation of the soluble th ...
Rattus norvegicus
Med. Biol.
59
177-184
1981
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
2
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210013
Barchi
Membrane-associated thiamin tr ...
Rattus norvegicus
J. Biol. Chem.
251
193-197
1976
-
-
-
-
-
-
1
-
1
3
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
3
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210014
Barchi
Membrane thiamine triphosphata ...
Rattus norvegicus
J. Neurochem.
26
715-720
1976
-
-
-
-
-
-
4
-
2
-
-
-
-
2
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
2
-
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210011
Iwata
Properties of thiamine di- and ...
Rattus norvegicus
J. Neurochem.
24
1209-1213
1975
-
-
-
-
-
-
4
-
2
-
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
2
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210009
Iwata
Role of thiamine metabolism in ...
Rattus norvegicus
Jpn. J. Pharmacol.
24
825-829
1974
-
-
-
-
-
-
2
-
1
-
-
-
-
2
-
-
-
-
-
3
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
3
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210010
Iwata
Role of thiamine metabolism in ...
Rattus norvegicus
Jpn. J. Pharmacol.
24
817-823
1974
-
-
-
-
-
-
1
-
3
2
-
-
-
2
-
-
-
-
-
3
-
-
4
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
3
2
-
-
-
-
-
-
-
3
-
-
4
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
210015
Iwata
-
Some properties of the enzyme ...
Rattus norvegicus
Thiamine (Proc. Pap. U. S. -Jpn. Semin, 2. Meeting)
2. Meeting
213-221
1974
-
-
-
-
-
-
4
-
2
2
-
-
-
1
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
2
2
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210016
Barchi
-
Thiamine triphosphatases in br ...
Rattus norvegicus
Thiamine (Proc. Pap. U. S. -Jpn. Semin, 2. Meeting)
195-212
1974
-
-
-
-
-
-
5
2
-
3
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
1
-
1
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
5
-
2
-
3
-
-
-
-
-
-
-
1
-
-
1
-
1
-
1
-
2
-
-
-
-
-
-
-
-
-
210006
Barchi
A membrane-associated thiamine ...
Rattus norvegicus
J. Biol. Chem.
247
7668-7673
1972
-
-
-
-
-
-
3
3
1
8
1
-
-
2
-
-
1
-
-
8
-
-
1
-
-
-
-
1
-
2
2
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
3
1
8
1
-
-
-
-
1
-
8
-
-
1
-
-
-
1
-
2
2
-
-
-
-
-
-
-
-
210007
Hashitani
The partial purification of th ...
Rattus norvegicus
J. Biol. Chem.
247
2117-2119
1972
-
-
-
-
-
-
4
1
1
-
-
-
-
2
-
-
-
-
-
6
1
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
1
1
-
-
-
-
-
-
-
-
6
1
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
210008
Barchi
Thiamine triphosphatase in the ...
Rattus norvegicus
Biochim. Biophys. Acta
255
402-405
1972
1
-
-
-
-
-
1
-
2
-
-
-
-
1
-
-
-
-
-
1
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
1
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-