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 (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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
gene PTPN22 | - |
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 | Comment | Organism |
---|---|---|
lymphoid-specific tyrosine phosphatase | - |
Homo sapiens |
LyP | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
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 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 |