High-Throughput Functional Screening of Steroid Substrates with Wild-Type and Chimeric P450 Enzymes
Panel (a), MDS configuration plot of the 31 CYP1A enzymes tested against all steroid activities. Some chimeric CYP1A enzymes are hidden replicates; that is, the cluster named “1” contains three chimeras of the Chim series which are triplicate of the same chimeric sequence. Solid circles correspond to wild-type enzymes, grey-filled circles to bi- or tripartite chimeras (Chim series), and open circles to mosaic chimeras (ChiMo series). The red ellipses correspond to cluster of enzymes which are discussed text based on their highly similar behaviour towards the steroids tested in this work. Panel (b), phylofunctional analysis of mouse-rabbit CYP1A chimeric enzymes assayed for hydroxylase activity with sixteen steroidal substrates. The open and solid bars represent, respectively, mouse CYP1A1 and rabbit CYP1A2 amino acid sequence segments. The purple and salmon boxes highlight the two elements of primary structure which are found to be correlated with the groups on the MDS plot.