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Literature summary for 3.1.3.48 extracted from

  • Yu, X.; Chen, M.; Zhang, S.; Yu, Z.H.; Sun, J.P.; Wang, L.; Liu, S.; Imasaki, T.; Takagi, Y.; Zhang, Z.Y.
    Substrate specificity of lymphoid-specific tyrosine phosphatase (Lyp) and identification of Src kinase-associated protein of 55 kDa homolog (SKAP-HOM) as a Lyp substrate (2011), J. Biol. Chem., 286, 30526-30534.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of N-terminally His-tagged enzyme mutant C227S, expression of Myc-tagged lymphoid-specific tyrosine phosphatase in HEK-293 cells, co-expression with SKAP-HOM, a cytosolic adaptor protein required for proper activation of the immune system, a bona fide Lyp substrate Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
recombinant N-terminally His-tagged inactive enzyme mutant C227S, im complex with the consensus peptide substrate or SKAP-HOM, a bona fide Lyp substrate, 3-5 days, X-ray diffraction structure determmination and analysis at 2.5 A and 2.9 A resolution, respectively, molecular replacement Homo sapiens

Protein Variants

Protein Variants Comment Organism
C195A site-directed mutagenesis, catalytically inactive mutant comprising residues 1-294, specific interaction between the Lyp substrate-trapping mutant and SKAP-HOM Homo sapiens
C227S site-directed mutagenesis, catalytically inactive mutant comprising residues 1-294, specific interaction between the Lyp substrate-trapping mutant and SKAP-HOM Homo sapiens
additional information GST pull-down experiment with pervanadate-treated, to increase and preserve tyrosine phosphorylation RAW264.7 cell lysates with either GST alone, wild-type Lyp fused to GST (GST-Lyp), or the substrate-trapping fusion proteins GST-Lyp/C227S and GST-Lyp/D195A. Whereas no protein retained by GST or GST-Lyp is detectable, the GST-Lyp/C227S and GST-Lyp/D195A trapping mutants specifically bound SKAP-HOM, the failure to detect association of SKAP-HOM with wild-type Lyp indicates that the interaction between the Lyp trapping mutants and SKAP-HOM requires tyrosine phosphorylation Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
gene PTPN22
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Ac-AAAApYAAAA-NH2 + H2O
-
Homo sapiens Ac-AAAAYAAAA-NH2 + phosphate
-
?
Ac-AAAEpYAAAA-NH2 + H2O
-
Homo sapiens Ac-AAAEYAAAA-NH2 + phosphate
-
?
Ac-AAAQpYAAAA-NH2 + H2O
-
Homo sapiens Ac-AAAQYAAAA-NH2 + phosphate
-
?
Ac-DGEEpYDDPF-NH2 + H2O SKAP-HOM Tyr75 peptide Homo sapiens Ac-DGEEYDDPF-NH2 + phosphate
-
?
Ac-ENDEpYTARE-NH2 + H2O Lck Tyr394 peptide Homo sapiens Ac-ENDEYTARE-NH2 + phosphate
-
?
Ac-TEPQpYQPGE-NH2 + H2O Lck Tyr505 peptide Homo sapiens Ac-TEPQYQPGE-NH2 + phosphate
-
?
Ac-YGEEpYDDLY-NH2 + H2O consensus peptide 1 Homo sapiens Ac-YGEEYDDLY-NH2 + phosphate
-
?
Ac-YGYEpYDDEY-NH2 + H2O consensus peptide 2 Homo sapiens Ac-YGYEYDDEY-NH2 + phosphate
-
?
additional information substrate specificity of lymphoid-specific tyrosine phosphatase, identification of consensus sequence motifs for Lyp substrate recognition using an inverse alanine scanning combinatorial library approach, molecular determinants and molecular basis for Lyp substrate recognition, crystal structure analysis, overview Homo sapiens ?
-
?
SKAP-HOM + H2O a cytosolic adaptor protein required for proper activation of the immune system, a bona fide Lyp substrate Homo sapiens ?
-
?

Synonyms

Synonyms Comment Organism
lymphoid-specific tyrosine phosphatase
-
Homo sapiens
LyP
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Homo sapiens

General Information

General Information Comment Organism
additional information GST pull-down experiment with pervanadate-treated, to increase and preserve tyrosine phosphorylation, RAW264.7 cell lysates with either GST alone, wild-type Lyp fused to GST (GST-Lyp), or the substrate-trapping fusion proteins GST-Lyp/C227S and GST-Lyp/D195A. Whereas no protein retained by GST or GST-Lyp is detectable, the GST-Lyp/C227S and GST-Lyp/D195A trapping mutants specifically bound SKAP-HOM, the failure to detect association of SKAP-HOM with wild-type Lyp indicates that the interaction between the Lyp trapping mutants and SKAP-HOM requires tyrosine phosphorylation Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00005
-
Ac-AAAApYAAAA-NH2 pH 7.0, 25°C Homo sapiens
0.0003
-
Ac-AAAQpYAAAA-NH2 pH 7.0, 25°C Homo sapiens
0.0006
-
Ac-TEPQpYQPGE-NH2 pH 7.0, 25°C Homo sapiens
0.0016
-
Ac-AAAEpYAAAA-NH2 pH 7.0, 25°C Homo sapiens
0.0062
-
Ac-ENDEpYTARE-NH2 pH 7.0, 25°C Homo sapiens
0.021
-
Ac-YGYEpYDDEY-NH2 pH 7.0, 25°C Homo sapiens
0.04
-
Ac-DGEEpYDDPF-NH2 pH 7.0, 25°C Homo sapiens
0.046
-
Ac-YGEEpYDDLY-NH2 pH 7.0, 25°C Homo sapiens