Membrane cholesterol as regulator of human rhomboid protease rhbdl4

HIGHLIGHTS

• who: Sandra Paschkowsky from the DNA constructs Plasmid p, containing cDNA encoding for human , with a, terminal Myc-FLAG tag was obtained from OriGeneInactive , S A was generated by site-directed mutagenesis using the forward primer, ⬘-GCTGTAGGTTTCGCAGGAGTTTTGTTT-3⬘ and the reverse primer, ⬘-AAACAAAACTCCTGCGAAACCTACAGC-3⬘. Mutants deficient in cholesterol binding, namely , Y A, Y A, and , / A, were cloned using gene blocks from IDT. Specifically, gene blocks were dissolved in H O at a concentration of , ng/␮l, of which , ng/␮l DNA were digested using XbaI and XmaI restriction enzymes. Enzymes were subsequently heat-inactivated at , °C . . .
  

   

  

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