Recombinant CYP3A4 Variants Production

Human full-length and Δ3-22 CYP3A4, as well as the R212A and T224A variants were produced as reported previously (21 (link), 39 (link)). The F219A mutation was introduced using PrimeSTAR Max DNA Polymerase master mix (Takara Bio) and confirmed by sequencing. The F219A variant of Δ3-22 CYP3A4 was expressed and purified using the protocol developed for WT CYP3A4 (39 (link)). All proteins used in the study had R_z (A_{416/280 nm} ratio) of 1.5 or higher.

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, & Sevrioukova I.F. (2023). Interaction of CYP3A4 with caffeine: First insights into multiple substrate binding. The Journal of Biological Chemistry, 299(9), 105117.

Publication 2023

PrimeSTAR Max DNA Polymerase master mix

Cyp3a4 Dna polymerase Human Mutation Proteins

Corresponding Organization : University of California, Irvine

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Variable analysis

independent variables

Human full-length CYP3A4
Human Δ3-22 CYP3A4
R212A variant
T224A variant
F219A variant of Δ3-22 CYP3A4

dependent variables

Purity of proteins (R_z ratio)

control variables

Purification protocol developed for wild-type CYP3A4

controls

Positive control: Wild-type CYP3A4

Annotations

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