Post on 02-May-2018
Cell, Volume 129
Supplemental Data
Structural Basis for the Inhibition of
Tyrosine Kinase Activity of ZAP-70 Sebastian Deindl, Theresa A. Kadlecek, Tomas Brdicka, Xiaoxian Cao, Arthur Weiss, and John Kuriyan
Table S1. Data Collection, Phasing, and Refinement Statistics
Data Collectiona
Space Group Unit Cell
# Molecules/asymmetric unit X-ray source Wavelength (Å) Resolution (Å) I/
Completeness (%) Rsym (%)
P1 a = 48.1 Å, b = 52.8 Å, c = 68.8 Å,
= 106.0°, = 93.1°, = 103.8° 1 Advanced Light Source, Berkeley, beamline 8.2.2 0.9537 50-2.6 16.2 (2.1) 95.3 (78.5) 8.7 (33.3)
Model Refinementb
Resolution (Å) # Reflections Rwork/Rfree
Rwork/Rfree (%) # Non-hydrogen protein/ligand atoms # Water molecules Rmsd bond length (Å) Rmsd angles (°) Average temperature (B) factor, protein (Å2)
50-2.6 15680/1622 22.0/29.0 4224/31 69 0.008 1.460 42.8
aValues as defined in SCALEPACK (Otwinowski and Minor, 1997); numbers in parentheses are statistics for the outer resolution shell (2.69-2.6 Å). bValues as defined in CNS (Brunger et al., 1998).
Figure S1. Conformers of the SH2-kinase linker.
Two alternate conformations for the SH2-kinase linker segment spanning residues 313-
322, as obtained from crystallographic simulated annealing twin refinements. The two
different conformers are colored green and blue.
Figure S2. Distance difference matrices.
Left: Structure of the kinase domain of ZAP-70 in the inactive Src/CDK-like
conformation. Regions that undergo structural changes upon adopting the active
conformation are colored. Right: Distance difference matrix between ZAP-70 in the
inactive Src/CDK-like conformation and ZAP-70 in the active conformation (PDB code:
1U59). Each entry in this symmetrical matrix represents the difference in the distance
between the corresponding C
atoms. C
atoms that come closer together on going from
the inactive Src/CDK-like conformation to the active conformation are indicated in
yellow/red. Distances that do not change much are green, and distances that expand are
blue. Data were cut off to +/- 5 Å.
Figure S3. Sequence alignment of ZAP-70 and Syk from different species, human
Src, human EphB2 and human Jak2.
Sequence alignment of ZAP-70/Syk with other tyrosine kinases. Secondary structure
elements (only for the kinase domain) are derived from the crystal structure of inactive
ZAP-70. Alignment was performed with ClustalW and formatted with ESPript using the
web interface.
Figure S4. Mutagenesis analysis of the interface.
LAT phosphorylation by diverse ZAP-70 mutants in the absence (left) or presence (right)
of Lck.
Figure S5.
The effect of hotspot mutations on ZAP-70 Y315F/Y319F dependent LAT
phosphorylation in the absence (A) or presence (B) of Lck.