Matches in SemOpenAlex for { <https://semopenalex.org/work/W2024174597> ?p ?o ?g. }
- W2024174597 endingPage "15761" @default.
- W2024174597 startingPage "15753" @default.
- W2024174597 abstract "The ZAP-70 protein tyrosine kinase is essential for T cell antigen receptor (TCR)-mediated signaling. The absence of ZAP-70 results in impaired differentiation of T cells and a lack of responsiveness to antigenic stimulation. In order to study the characteristics of ZAP-70 in vitro, we overexpressed an epitopically tagged human ZAP-70 in a recombinant baculovirus expression system and purified it by column chromatography. The kinase activity of purified, recombinant ZAP-70 required cation and exhibited a strong preference for Mn2+ over Mg2+. The apparent Km of ZAP-70 for ATP was ∼3.0 µM. The activity of the recombinant ZAP-70, unlike that of the homologous protein tyrosine kinase, Syk, was not affected by binding of TCR-derived tyrosine phosphorylated immunoreceptor tyrosine-based activation motif peptides. Several proteins were tested as potential in vitro substrates of ZAP-70. Only α-tubulin and the cytoplasmic fragment of human erythrocyte band 3 (cfb3), which have a region of sequence identity at the phosphorylation site, proved to be good substrates, exhibiting Km values of ∼3.3 and ∼2.5 µM, respectively ([ATP] = 50 µM). α- and β-Casein were poor substrates for ZAP-70, and no activity toward enolase, myelin basic protein, calmodulin, histone proteins, or angiotensin could be detected. In contrast to the T cell protein tyrosine kinase, Lck, ZAP-70 did not phosphorylate the cytoplasmic portion of the TCRζ chain or short peptides corresponding to the CD3ε or the TCRζ immunoreceptor tyrosine-based activation motifs. Our studies suggest that ZAP-70 exhibits a high degree of substrate specificity. The ZAP-70 protein tyrosine kinase is essential for T cell antigen receptor (TCR)-mediated signaling. The absence of ZAP-70 results in impaired differentiation of T cells and a lack of responsiveness to antigenic stimulation. In order to study the characteristics of ZAP-70 in vitro, we overexpressed an epitopically tagged human ZAP-70 in a recombinant baculovirus expression system and purified it by column chromatography. The kinase activity of purified, recombinant ZAP-70 required cation and exhibited a strong preference for Mn2+ over Mg2+. The apparent Km of ZAP-70 for ATP was ∼3.0 µM. The activity of the recombinant ZAP-70, unlike that of the homologous protein tyrosine kinase, Syk, was not affected by binding of TCR-derived tyrosine phosphorylated immunoreceptor tyrosine-based activation motif peptides. Several proteins were tested as potential in vitro substrates of ZAP-70. Only α-tubulin and the cytoplasmic fragment of human erythrocyte band 3 (cfb3), which have a region of sequence identity at the phosphorylation site, proved to be good substrates, exhibiting Km values of ∼3.3 and ∼2.5 µM, respectively ([ATP] = 50 µM). α- and β-Casein were poor substrates for ZAP-70, and no activity toward enolase, myelin basic protein, calmodulin, histone proteins, or angiotensin could be detected. In contrast to the T cell protein tyrosine kinase, Lck, ZAP-70 did not phosphorylate the cytoplasmic portion of the TCRζ chain or short peptides corresponding to the CD3ε or the TCRζ immunoreceptor tyrosine-based activation motifs. Our studies suggest that ZAP-70 exhibits a high degree of substrate specificity." @default.
- W2024174597 created "2016-06-24" @default.
- W2024174597 creator A5006158729 @default.
- W2024174597 creator A5028727045 @default.
- W2024174597 creator A5053361384 @default.
- W2024174597 creator A5058111648 @default.
- W2024174597 creator A5089066152 @default.
- W2024174597 date "1996-06-01" @default.
- W2024174597 modified "2023-10-12" @default.
- W2024174597 title "Purification and Characterization of Human ZAP-70 Protein-tyrosine Kinase from a Baculovirus Expression System" @default.
- W2024174597 cites W133456552 @default.
- W2024174597 cites W1489480230 @default.
- W2024174597 cites W1495954528 @default.
- W2024174597 cites W1503069128 @default.
- W2024174597 cites W1516065974 @default.
- W2024174597 cites W1518448964 @default.
- W2024174597 cites W1542980948 @default.
- W2024174597 cites W1547466369 @default.
- W2024174597 cites W1549736588 @default.
- W2024174597 cites W1568846447 @default.
- W2024174597 cites W1575863011 @default.
- W2024174597 cites W1577765297 @default.
- W2024174597 cites W1581148997 @default.
- W2024174597 cites W1583957430 @default.
- W2024174597 cites W1587792888 @default.
- W2024174597 cites W1608816599 @default.
- W2024174597 cites W1649862870 @default.
- W2024174597 cites W1963569988 @default.
- W2024174597 cites W1965703662 @default.
- W2024174597 cites W1975085160 @default.
- W2024174597 cites W1983960385 @default.
- W2024174597 cites W1996417579 @default.
- W2024174597 cites W2004458954 @default.
- W2024174597 cites W2006363406 @default.
- W2024174597 cites W2007794993 @default.
- W2024174597 cites W2009597900 @default.
- W2024174597 cites W2012694659 @default.
- W2024174597 cites W2017037387 @default.
- W2024174597 cites W2019670504 @default.
- W2024174597 cites W2023090301 @default.
- W2024174597 cites W2026907934 @default.
- W2024174597 cites W2028847269 @default.
- W2024174597 cites W2030348245 @default.
- W2024174597 cites W2041255103 @default.
- W2024174597 cites W2042209841 @default.
- W2024174597 cites W2042582068 @default.
- W2024174597 cites W2044219128 @default.
- W2024174597 cites W2044336492 @default.
- W2024174597 cites W2052561585 @default.
- W2024174597 cites W2055444005 @default.
- W2024174597 cites W2058441584 @default.
- W2024174597 cites W2065279599 @default.
- W2024174597 cites W2067884008 @default.
- W2024174597 cites W2068982799 @default.
- W2024174597 cites W2074059194 @default.
- W2024174597 cites W2074416628 @default.
- W2024174597 cites W2079003536 @default.
- W2024174597 cites W2083319707 @default.
- W2024174597 cites W2087983099 @default.
- W2024174597 cites W2094126922 @default.
- W2024174597 cites W2095433140 @default.
- W2024174597 cites W2099661341 @default.
- W2024174597 cites W2131894799 @default.
- W2024174597 cites W2134020845 @default.
- W2024174597 cites W2144935474 @default.
- W2024174597 cites W2147753962 @default.
- W2024174597 cites W2151151593 @default.
- W2024174597 cites W2184963582 @default.
- W2024174597 cites W2186444379 @default.
- W2024174597 cites W4293247451 @default.
- W2024174597 cites W78775163 @default.
- W2024174597 doi "https://doi.org/10.1074/jbc.271.26.15753" @default.
- W2024174597 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/8663155" @default.
- W2024174597 hasPublicationYear "1996" @default.
- W2024174597 type Work @default.
- W2024174597 sameAs 2024174597 @default.
- W2024174597 citedByCount "55" @default.
- W2024174597 countsByYear W20241745972012 @default.
- W2024174597 countsByYear W20241745972014 @default.
- W2024174597 countsByYear W20241745972016 @default.
- W2024174597 countsByYear W20241745972018 @default.
- W2024174597 countsByYear W20241745972021 @default.
- W2024174597 countsByYear W20241745972022 @default.
- W2024174597 crossrefType "journal-article" @default.
- W2024174597 hasAuthorship W2024174597A5006158729 @default.
- W2024174597 hasAuthorship W2024174597A5028727045 @default.
- W2024174597 hasAuthorship W2024174597A5053361384 @default.
- W2024174597 hasAuthorship W2024174597A5058111648 @default.
- W2024174597 hasAuthorship W2024174597A5089066152 @default.
- W2024174597 hasBestOaLocation W20241745971 @default.
- W2024174597 hasConcept C108636557 @default.
- W2024174597 hasConcept C11960822 @default.
- W2024174597 hasConcept C153911025 @default.
- W2024174597 hasConcept C19317047 @default.
- W2024174597 hasConcept C196347352 @default.
- W2024174597 hasConcept C197153747 @default.
- W2024174597 hasConcept C203014093 @default.
- W2024174597 hasConcept C2776090121 @default.