| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | For Y2H, human profilaggrin N-terminal constructs containing the A (S100) domain, B domain or A domain plus varying amounts of the B domain were prepared using PCR and DNA cloning into the pOBD and pOAD vectors (Yeast Resource Center, University of Washington) (Hudson et al., 1997) (Table S5). For crystallization, PF-CABD (A domain, human profilaggrin residues 1-92 inclusive of N-terminal methionine) was purchased from GenScript (Piscataway, NJ) (transformed into plasmid pGS-21a) containing N-terminal 6x-histidine tag with thrombin cleavage site. | Get A Quote |
The fused-type S100 protein profilaggrin and its proteolytic products including filaggrin are important in the formation of a normal epidermal barrier; however, the specific function of the S100 calcium-binding domain in profilaggrin biology is poorly understood. To explore its molecular function, we determined a 2.2 Å-resolution crystal structure of the N-terminal fused-type S100 domain of human profilaggrin with bound calcium ions. The profilaggrin S100 domain formed a stable dimer, which contained two hydrophobic pockets that provide a molecular interface for protein interactions. Biochemical and molecular approaches demonstrated that three proteins, annexin II/p36, stratifin/14-3-3 sigma, and heat shock ... More
