Journal: The Journal of Biological Chemistry

Article Title: Disease-causing cystathionine β-synthase linker mutations impair allosteric regulation

doi: 10.1016/j.jbc.2023.105449

Figure Lengend Snippet: Comparison of L-serine-dependent cystathionine and cysteine-dependent H 2 S synthesis specific activities by wild-type and linker variants of human CBS a

Article Snippet: Human CBS carrying a 10 amino acid deletion (residues 516–525) in the regulatory domain was purchased from GenScript and cloned into the pET28b+ vector incorporating a cleavable C-terminal 6-His tag.

Techniques: Comparison

Journal: The Journal of Biological Chemistry

Article Title: Disease-causing cystathionine β-synthase linker mutations impair allosteric regulation

doi: 10.1016/j.jbc.2023.105449

Figure Lengend Snippet: Structures of dimeric human CBS in the basal and activated conformations. A , scheme showing the intersection between the methionine cycle ( grey ) and the transsulfuration pathway ( black ). For clarity, only the dominant H 2 S-generating reactions catalyzed by CBS and CTH are shown in red . α-KB is α-ketobutyrate. B , modular organization of CBS. C and D , the basal (PDB: 4COO , C ) and activated (PDB: 4PCU , D ) conformations of the CBS Δ516–525 dimer are shown using the same color scheme as in B . The two subunits are shown in dark and light shades. Heme, PLP, and AdoMet (in the activated conformation) are shown in red , yellow , and cyan spheres , respectively. E , schematic representation of the large conformational changes seen in the crystal structure of the CBS Δ516–525 dimer and the cryo-EM structure of the native fibrillar form. AdoMet either triggers or stabilizes the activated conformation in which the regulatory domains dimerize atop the catalytic cores, dramatically changing the organization of the fibrillar form.

Article Snippet: Human CBS carrying a 10 amino acid deletion (residues 516–525) in the regulatory domain was purchased from GenScript and cloned into the pET28b+ vector incorporating a cleavable C-terminal 6-His tag.

Techniques: Cryo-EM Sample Prep