Figure 3. Furin cleavage does not affect the structure of the human Notch2 HD domain.

(A) ¹⁵N HSQC spectrum of uniformly ¹⁵N-labeled human Notch2 HD domain before (black, left panel) and after (red, center panel) in vitro furin processing. A superposition of the two spectra is also shown (right panel). After acquisition of the initial spectrum of the human Notch2 HD domain (350 mM) at 18°C in 50 mM Bis-Tris (pH 7.0) containing 1 mM CaCl₂, 50 mM NaCl, the protein was incubated for 13.5 hrs with 150 µl furin at 37°C. A second spectrum was then taken at 18°C. (B) HSQC spectrum of the human Notch2 HD domain selectively labeled with ¹⁵N-Leucine before (black, left panel) and after (red, center panel) furin cleavage. A superposition of the two spectra is also shown (right panel). Spectra in (B) were acquired at pH 6.0; all other conditions were identical to (A).