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Literature summary for 3.4.14.5 extracted from

  • Aertgeerts, K.; Ye, S.; Shi, L.; Prasad, S.G.; Witmer, D.; Chi, E.; Sang, B.C.; Wijnands, R.A.; Webb, D.R.; Swanson, R.V.
    N-linked glycosylation of dipeptidyl peptidase IV (CD26): effects on enzyme activity, homodimer formation, and adenosine deaminase binding (2004), Protein Sci., 13, 145-154.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.14.5

Cloned(Commentary)

Cloned (Comment) Organism
expression of a soluble His-tagged wild-type enzyme and glycosylation mutant extracellular domains, which comprises extracellular domain amino acid residues 39-766 and are secreted to the medium, in Trichoplusia ni Hi5 and Spodoptera frugiperda Sf9 insect cells via baculovirus infection Homo sapiens
expression of wild-type enzyme and mutant enzymes in Sf9 insect cells and in Hi5 insect cells Homo sapiens

Protein Variants

Protein Variants Comment Organism
N150A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, 60% increased activity if expressed in Hi5 cells, unaltered activity if expressed in Sf9 cells Homo sapiens
N150A kcat/Km for mutant enzyme expressed in Sf9 insect cells is identical to of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 110% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens
N219A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, increased binding to adenosine deaminase of the mutant if expressed in Sf9 cells, increased activity if expressed in Hi5 cells, slightly altered activity if expressed in Sf9 cells Homo sapiens
N219A kcat/Km for mutant enzyme expressed in Sf9 insect cells is 92% of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 110% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens
N229A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, 90% increased activity if expressed in Hi5 cells, reduced activity if expressed in Sf9 cells Homo sapiens
N229A kcat/Km for mutant enzyme expressed in Sf9 insect cells is 70% of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 73% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens
N281A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, 40% increased activity if expressed in Hi5 cells, reduced activity if expressed in Sf9 cells Homo sapiens
N281A kcat/Km for mutant enzyme expressed in Sf9 insect cells is 85% of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 92% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens
N321A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, slightly increased binding to adenosine deaminase of the mutant if expressed in Sf9 cells, increased activity if expressed in Hi5 cells, reduced activity if expressed in Sf9 cells Homo sapiens
N321A kcat/Km for mutant enzyme expressed in Sf9 insect cells is 50% of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 54% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens
N520A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, slightly increased binding to adenosine deaminase of the mutant if expressed in Sf9 cells Homo sapiens
N520A kcat/Km for mutant enzyme expressed in Sf9 insect cells is 68% of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 80% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens
N685A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, slightly decreased binding to adenosine deaminase of the mutant if expressed in Hi5 cells Homo sapiens
N685A kcat/Km for mutant enzyme expressed in Sf9 insect cells is 110% of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 120% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens
N85A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, slightly increased binding to adenosine deaminase of the mutant if expressed in Sf9 cells, increased activity if expressed in Hi5 cells, slightlyreduced activity if expressed in Sf9 cells Homo sapiens
N85A kcat/Km for mutant enzyme expressed in Sf9 insect cells is 85% of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 92% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens
N92A site-directed mutagenesis, glycosylation site mutant, unaltered dimerization compared to the wild-type, slightly increased activity if expressed in Hi5 cells, unaltered activity if expressed in Sf9 cells Homo sapiens
N92A kcat/Km for mutant enzyme expressed in Sf9 insect cells is identical to of wild-type value for the substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin and 110% of wild-type value for the substrate Gly-Pro-4-nitroanilide Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.013
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N685A, expressed in Sf9 cells Homo sapiens
0.013
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N685A expressed in Sf9 cells Homo sapiens
0.014
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, wild-type soluble enzyme, mutant N92A, N150A, N229A, N281A, N321A and N520A, expressed in Sf9 cells Homo sapiens
0.014
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N150A expressed in Sf9 cells Homo sapiens
0.014
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N229A expressed in Sf9 cells Homo sapiens
0.014
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N281A expressed in Sf9 cells Homo sapiens
0.014
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N321A expressed in Sf9 cells Homo sapiens
0.014
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N520A expressed in Sf9 cells Homo sapiens
0.014
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N92A expressed in Sf9 cells Homo sapiens
0.014
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, wild-type enzyme expressed in Sf9 cells Homo sapiens
0.015
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N229A and N685A, expressed in Hi5 cells Homo sapiens
0.015
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N85A and N219A, expressed in Sf9 cells Homo sapiens
0.015
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N219A expressed in Sf9 cells Homo sapiens
0.015
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N85A expressed in Sf9 cells Homo sapiens
0.016
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, wild-type soluble enzyme and mutant N85A, expressed in Hi5 cells Homo sapiens
0.017
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N92A, N150A, and N520A, expressed in Hi5 cells Homo sapiens
0.018
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N321A, expressed in Hi5 cells Homo sapiens
0.019
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N219A, expressed in Hi5 cells Homo sapiens
0.02
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N281A, expressed in Hi5 cells Homo sapiens
0.131
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N520A, expressed in Hi5 cells Homo sapiens
0.138
-
 Gly-Pro-4-nitroanilide pH 7.4, wild-type soluble enzyme, expressed in Hi5 cells Homo sapiens
0.142
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N520A, expressed in Sf9 cells Homo sapiens
0.142
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N685A, expressed in Hi5 cells Homo sapiens
0.143
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N150A, expressed in Sf9 cells Homo sapiens
0.143
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N92A, expressed in Hi5 cells Homo sapiens
0.144
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N219A, expressed in Sf9 cells Homo sapiens
0.145
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N229A, expressed in Hi5 cells Homo sapiens
0.146
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N321A, expressed in Hi5 cells Homo sapiens
0.147
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N92A, expressed in Sf9 cells Homo sapiens
0.149
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N85A, expressed in Hi5 cells Homo sapiens
0.151
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N150A, expressed in Hi5 cells Homo sapiens
0.151
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N229A, expressed in Sf9 cells Homo sapiens
0.152
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N685A, expressed in Sf9 cells Homo sapiens
0.155
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N85A, expressed in Sf9 cells Homo sapiens
0.156
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N321A, expressed in Sf9 cells Homo sapiens
0.159
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N281A, expressed in Sf9 cells Homo sapiens
0.16
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N219A, expressed in Hi5 cells Homo sapiens
0.168
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N281A, expressed in Hi5 cells Homo sapiens
0.17
-
 Gly-Pro-4-nitroanilide pH 7.4, wild-type soluble enzyme, expressed in Sf9 cells Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane type II transmembrane protein Homo sapiens 16020
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
95000
-
 2 * 95000, SDS-PAGE Homo sapiens
220000
-
 gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Homo sapiens binding of adenosine deaminase to the T-cell surface protects the T-cell from adenosine-mediated inhibition of proliferation ?
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein 9 N-glycosylation sites, glycosylation is no prerequisite for enzyme activity, dimerization or binding of adenosine deaminase, structural analysis Homo sapiens

Purification (Commentary)

Purification (Comment) Organism
recombinant soluble His-tagged wild-type and mutant extracellular enzyme domains from insect cell culture supernatant to homogeneity Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Ala-Pro-7-amido-4-methylcoumarin + H2O
-
 Homo sapiens Ala-Pro + 7-amino-4-methylcoumarin
-
 ?
Ala-Pro-7-amido-4-trifluoromethylcoumarin + H2O
-
 Homo sapiens Ala-Pro + 7-amino-4-trifluoromethylcoumarin
-
 ?
Gly-Pro-4-nitroanilide + H2O
-
 Homo sapiens Gly-Pro + 4-nitroaniline
-
 ?
additional information enzyme binds adenosine deaminase at the highly glycosylated beta-propeller domain Homo sapiens ?
-
 ?
additional information binding of adenosine deaminase to the T-cell surface protects the T-cell from adenosine-mediated inhibition of proliferation Homo sapiens ?
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 95000, SDS-PAGE Homo sapiens

Synonyms

Synonyms Comment Organism
ADABP
-
 Homo sapiens
adenosine deaminase binding protein
-
 Homo sapiens
CD26
-
 Homo sapiens
dipeptidyl peptidase IV
-
 Homo sapiens
DPPIV
-
 Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3 6 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N520A, expressed in Hi5 cells Homo sapiens
9
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N321A, expressed in Sf9 cells Homo sapiens
9
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N321A expressed in Sf9 cells Homo sapiens
10
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N321A, expressed in Sf9 cells Homo sapiens
12
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N229A, expressed in Sf9 cells Homo sapiens
12
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N229A expressed in Sf9 cells Homo sapiens
13
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N229A, expressed in Sf9 cells Homo sapiens
13
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N520A, expressed in Sf9 cells Homo sapiens
13
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N520A expressed in Sf9 cells Homo sapiens
14
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N520A, expressed in Sf9 cells Homo sapiens
15
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N281A, expressed in Sf9 cells Homo sapiens
15
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N281A expressed in Sf9 cells Homo sapiens
16
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N85A, expressed in Sf9 cells Homo sapiens
16
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N85A, expressed in Sf9 cells Homo sapiens
16
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N85A expressed in Sf9 cells Homo sapiens
17
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N281A, expressed in Sf9 cells Homo sapiens
18
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, wild-type soluble enzyme, mutant N150A, N219A, and N92A, expressed in Sf9 cells Homo sapiens
18
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N150A expressed in Sf9 cells Homo sapiens
18
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N219A expressed in Sf9 cells Homo sapiens
18
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N92A expressed in Sf9 cells Homo sapiens
18
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, wild-type enzyme expressed in Sf9 cells Homo sapiens
19
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N685A, expressed in Sf9 cells Homo sapiens
19
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N92A, N150A, and N219A, expressed in Sf9 cells Homo sapiens
19
-
 Ala-Pro-7-amido-4-trifluoromethylcoumarin pH 7.4, mutant enzyme N685A expressed in Sf9 cells Homo sapiens
20
-
 Gly-Pro-4-nitroanilide pH 7.4, wild-type soluble enzyme, expressed in Sf9 cells Homo sapiens
21
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N685A, expressed in Sf9 cells Homo sapiens
23
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, wild-type soluble enzyme, expressed in Hi5 cells Homo sapiens
24
-
 Gly-Pro-4-nitroanilide pH 7.4, wild-type soluble enzyme, expressed in Hi5 cells Homo sapiens
27
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N85A, expressed in Hi5 cells Homo sapiens
30
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N685A, expressed in Hi5 cells Homo sapiens
30
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N85A, expressed in Hi5 cells Homo sapiens
31
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N92A, expressed in Hi5 cells Homo sapiens
32
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N685A, expressed in Hi5 cells Homo sapiens
34
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N321A, expressed in Hi5 cells Homo sapiens
34
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N92A, expressed in Hi5 cells Homo sapiens
35
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N321A and N520A, expressed in Hi5 cells Homo sapiens
37
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N150A, expressed in Hi5 cells Homo sapiens
40
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N150A, expressed in Hi5 cells Homo sapiens
40
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N229A and N281A, expressed in Hi5 cells Homo sapiens
42
-
 Ala-Pro-7-amido-4-methylcoumarin pH 7.4, mutant N219A, expressed in Hi5 cells Homo sapiens
43
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N229A and N281A, expressed in Hi5 cells Homo sapiens
45
-
 Gly-Pro-4-nitroanilide pH 7.4, mutant N219A, expressed in Hi5 cells Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
 assay at Homo sapiens