Conformational States Control Lck Switching between Free and Confined Diffusion Modes in T Cells.

Abstract:

:T cell receptor phosphorylation by Lck is an essential step in T cell activation. It is known that the conformational states of Lck control enzymatic activity; however, the underlying principles of how Lck finds its substrate over the plasma membrane remain elusive. Here, single-particle tracking is paired with photoactivatable localization microscopy to observe the diffusive modes of Lck in the plasma membrane. Individual Lck molecules switched between free and confined diffusion in both resting and stimulated T cells. Lck mutants locked in the open conformation were more confined than Lck mutants in the closed conformation. Further confinement of kinase-dead versions of Lck suggests that Lck confinement was not caused by phosphorylated substrates. Our data support a model in which confined diffusion of open Lck results in high local phosphorylation rates, and inactive, closed Lck diffuses freely to enable long-range distribution over the plasma membrane.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Hilzenrat G,Pandžić E,Yang Z,Nieves DJ,Goyette J,Rossy J,Ma Y,Gaus K

doi

10.1016/j.bpj.2020.01.041

subject

Has Abstract

pub_date

2020-03-24 00:00:00

pages

1489-1501

issue

6

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(20)30116-8

journal_volume

118

pub_type

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