recombinant human ubiquitin protein  (Proteintech)

Journal: Biochemistry and Biophysics Reports

Article Title: Reconstitution of CFTR ubiquitination identifies lysine-420 as a regulator of cell surface residence and current

doi: 10.1016/j.bbrep.2025.102393

Figure Lengend Snippet: Ubiquitinated lysines identified in this study. Peptide sequences and corresponding amino acid numbers are shown, with ubiquitinated lysines highlighted in red. The analysis was performed twice, and ∗ denotes that a ubiquitin modification was isolated in both trials. CFTR peptide coverage was 54 % for experiment 1 and 60 % for experiment 2.

Article Snippet: Reactions contained 160 nM of His-UBE1 (R&D Systems #E−304; also known as UBA1), 830 nM of UbcH5b/UBE2D2 (R&D Systems #E2-622), 37 μM of human recombinant ubiquitin (R&D Systems #U–100H), 7 μM CHIP (an E3 ligase, purified as described in Ref. [ ]), and 1.3 μM of detergent-solubilized purified CFTR (see above) in an ATP regeneration buffer (1 mM ATP, 50 mM creatine phosphate, 200 μg/mL creatine phosphokinase, 20 mM HEPES, pH 6.8, 150 mM potassium acetate, 5 mM magnesium acetate, 250 mM sorbitol).

Techniques: Ubiquitin Proteomics, Modification, Isolation