title: T cell receptor dwell times control the kinase activity of Zap70.
abstract: Kinase recruitment to membrane receptors is essential for signal transduction. However, the underlying regulatory mechanisms are poorly understood. We investigated how conformational changes control T cell receptor (TCR) association and activity of the kinase Zap70. Structural analysis showed that TCR binding or phosphorylation of Zap70 triggers a transition from a closed, autoinhibited conformation to an open conformation. Using Zap70 mutants with defined conformations, we found that TCR dwell times controlled Zap70 activity. The closed conformation minimized TCR dwell times and thereby prevented activation by membrane-associated kinases. Parallel recruitment of coreceptor-associated Lck kinase to the TCR ensured Zap70 phosphorylation and stabilized Zap70 TCR binding. Our study suggests that the dynamics of cytosolic enzyme recruitment to the plasma membrane regulate the activity and function of receptors lacking intrinsic catalytic activity.
authors.affiliation: Nomis Center for Immunobiology and Microbial Pathogenesis &Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, California, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Klammt
authors.affiliation: Nomis Center for Immunobiology and Microbial Pathogenesis &Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, California, USA.
authors.forename: Lucie
authors.initials: L
authors.lastname: Novotná
authors.affiliation: Nomis Center for Immunobiology and Microbial Pathogenesis &Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, California, USA.
authors.forename: Dongyang T
authors.initials: DT
authors.lastname: Li
authors.affiliation: Nomis Center for Immunobiology and Microbial Pathogenesis &Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, California, USA.
authors.forename: Miriam
authors.initials: M
authors.lastname: Wolf
authors.affiliation: Nomis Center for Immunobiology and Microbial Pathogenesis &Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, California, USA.
authors.forename: Amy
authors.initials: A
authors.lastname: Blount
authors.affiliation: Eli Lilly Inc., Lilly Biotechnology Center, San Diego, California, USA.
authors.forename: Kai
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Eli Lilly Inc., Lilly Biotechnology Center, San Diego, California, USA.
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Fitchett
authors.affiliation: Nomis Center for Immunobiology and Microbial Pathogenesis &Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, California, USA.
authors.forename: Björn F
authors.initials: BF
authors.lastname: Lillemeier
mesh_terms: D017252:CD3 Complex / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D041961:Deuterium Exchange Measurement; D006801:Humans; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism*; D013058:Mass Spectrometry; D056004:Molecular Dynamics Simulation; D009154:Mutation; D010766:Phosphorylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000235:genetics / Q000378:metabolism*
chemical_list: D017252:CD3 Complex; D011948:Receptors, Antigen, T-Cell; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051746:ZAP-70 Protein-Tyrosine Kinase