Assuntos
Carcinoma de Células Escamosas/genética , Epidermólise Bolhosa Distrófica/genética , Proteínas Proto-Oncogênicas c-myc/genética , Receptores Notch/genética , Neoplasias Cutâneas/genética , Proteína Supressora de Tumor p53/genética , Adulto , Animais , Biópsia , Carcinoma de Células Escamosas/complicações , Carcinoma de Células Escamosas/patologia , Colágeno Tipo VII/genética , Variações do Número de Cópias de DNA , Epidermólise Bolhosa Distrófica/complicações , Epidermólise Bolhosa Distrófica/patologia , Feminino , Genes Recessivos/genética , Perfil Genético , Humanos , Perda de Heterozigosidade , Masculino , Camundongos , Neoplasias Cutâneas/complicações , Neoplasias Cutâneas/patologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Phosphorylation is a reversible post-translational modification, playing a vital role in protein function. In T cells, protein phosphorylation is the key mechanism regulating T cell receptor-driven signaling pathways. In order to gain insights into the phosphoproteome evolution of T cell activation, we performed a large-scale quantitative phosphoproteomics study of Jurkat E6.1 (wild type) and Jurkat gamma1 (Phospholipase gamma1 null) cell clones upon costimulation with anti-CD3 and anti-CD28 antibodies at times ranging from 15min to as long as 120min. In total, we identified 5585 phosphopeptides belonging to 2008 phosphoproteins from both cell clones. We detected 130 and 114 novel phosphopeptides in Jurkat E6.1 and Jurkat gamma1 clones, respectively. A significantly lower number of proteins containing regulated phosphorylation sites were identified in Jurkat gamma1 in comparison to Jurkat E6.1, reflecting the vital role of Phospholipase gamma1 in T cell signaling. Several new phosphorylation sites from lymphocyte-specific protein tyrosine kinase (Lck) were identified. Of these, serine-121 showed significant changes in JE6.1 while only small changes in the Jgamma1 clone. Our data may contribute to the current human T cell phosphoproteome and provide a better understanding on T cell receptor signaling. Data are available via ProteomeXchange with identifier PXD002871.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos CD28/imunologia , Complexo CD3/imunologia , Fosfoproteínas/imunologia , Proteoma/imunologia , Clonagem de Organismos , Humanos , Imunização , Células JurkatRESUMO
LymPHOS is a web-oriented database containing peptide and protein sequences and spectrometric information on the phosphoproteome of primary human T-Lymphocytes. Current release 2.0 contains 15 566 phosphorylation sites from 8273 unique phosphopeptides and 4937 proteins, which correspond to a 45-fold increase over the original database description. It now includes quantitative data on phosphorylation changes after time-dependent treatment with activators of the TCR-mediated signal transduction pathway. Sequence data quality has also been improved with the use of multiple search engines for database searching. LymPHOS can be publicly accessed at http://www.lymphos.org. Database URL: http://www.lymphos.org.
