Sham_control

Line scan images Ca transients in 12 fluo-3-AM loaded atrial myocytes from Sham-operated hearts stimulated at 1 Hz. Note: These are raw images; those presented in paper represent averages of consecutive transient

RyR_Sham-1

Images of 18 atrial myocytes stained with anti RyR-2 antibody from hearts from Sham-operated rat

sham 4Hz

Txt data from 12 fluo-3-AM loaded atrial myocytes from Sham operated rat hearts subject to 4 Hz pacing followed by a period of at least 70 s without stimulatio

Sham caffeine data

Line scan images of electrically stimulated and caffeine-induced Ca transients from 10 fluo-3-AM loaded atrial myocytes from Sham operated heart

Sham di-8-ANEPPS

Z-stacks of 20 di-8-ANEPPS-stained atrial myocytes from Sham operated rat heart

AoB_control

Line scan images of Ca transients from 8 fluo-3-AM loaded atrial myocytes from aortic banded rat hearts. Note: these are raw data of consecutive transients; transients presented in the paper represent averages of consecutive transient

AoB Hz

Txt data from 13 fluo-3-AM loaded atrial myocytes from aortic banded heart

AoB caffeine data

Line scan images of electrically stimulated and caffeine-induced Ca transients from 13 fluo-3-AM loaded atrial myocytes from aortic banded rat heart

Design and Nuclear Magnetic Resonance (NMR) Structure Determination of the Second Extracellular Immunoglobulin Tyrosine Kinase A (TrkAIg2) Domain Construct for Binding Site Elucidation in Drug Discovery

The tyrosine kinase A (TrkA) receptor is a validated therapeutic intervention point for a wide range of conditions. TrkA activation by nerve growth factor (NGF) binding the second extracellular immunoglobulin (TrkAIg2) domain triggers intracellular signaling cascades. In the periphery, this promotes the pain phenotype and, in the brain, cell survival or differentiation. Reproducible structural information and detailed validation of protein–ligand interactions aid drug discovery. However, the isolated TrkAIg2 domain crystallizes as a β-strand-swapped dimer in the absence of NGF, occluding the binding surface. Here we report the design and structural validation by nuclear magnetic resonance spectroscopy of the first stable, biologically active construct of the TrkAIg2 domain for binding site confirmation. Our structure closely mimics the wild-type fold of TrkAIg2 in complex with NGF (1WWW.pdb), and the ¹H–¹⁵N correlation spectra confirm that both NGF and a competing small molecule interact at the known binding interface in solution