5ZAU

Complex of the human FYN SH3 and monobody binder

Summary for 5ZAU

• Entry DOI: 10.2210/pdb5zau/pdb
• Descriptor: Tyrosine-protein kinase Fyn, Monobody Binder (2 entities in total)
• Functional Keywords: complex, signaling protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 13894.74

Authors

Reddy, P.P.,Gulyani, A.,Das, R. (deposition date: 2018-02-09, release date: 2019-09-11, Last modification date: 2024-05-15)

Primary citation

Mukherjee, A.,Singh, R.,Udayan, S.,Biswas, S.,Reddy, P.P.,Manmadhan, S.,George, G.,Kumar, S.,Das, R.,Rao, B.M.,Gulyani, A.
A Fyn biosensor reveals pulsatile, spatially localized kinase activity and signaling crosstalk in live mammalian cells.
Elife, 9:-, 2020

PubMed Abstract: Cell behavior is controlled through spatio-temporally localized protein activity. Despite unique and often contradictory roles played by Src-family-kinases (SFKs) in regulating cell physiology, activity patterns of individual SFKs have remained elusive. Here, we report a biosensor for specifically visualizing active conformation of SFK-Fyn in live cells. We deployed combinatorial library screening to isolate a binding-protein (F29) targeting activated Fyn. Nuclear-magnetic-resonance (NMR) analysis provides the structural basis of F29 specificity for Fyn over homologous SFKs. Using F29, we engineered a sensitive, minimally-perturbing fluorescence-resonance-energy-transfer (FRET) biosensor () that reveals cellular Fyn activity to be spatially localized, pulsatile and sensitive to adhesion/integrin signaling. Strikingly, growth factor stimulation further enhanced Fyn activity in pre-activated intracellular zones. However, inhibition of focal-adhesion-kinase activity not only attenuates Fyn activity, but abolishes growth-factor modulation. imaging uncovers spatially organized, sensitized signaling clusters, direct crosstalk between integrin and growth-factor-signaling, and clarifies how compartmentalized Src-kinase activity may drive cell fate.

PubMed: 32017701
DOI: 10.7554/eLife.50571

Experimental method

SOLUTION NMR