Figure 5
PGRMC1 expression plasmids expressing wild-type PGRMC1 and various amino acid mutants. Mutations were introduced by polymerase chain reaction, after which the entire open reading frame of product plasmids was confirmed correct by DNA sequencing. Individual codon mutations were encoded by the following sequences: serine-56 to alanine (S56A) AGC → GCC; cysteine-128 to serine (C128S) TGC → AGC; tyrosine-138 to phenylalanine (Y138F) TAC → TTC; tyrosine-179 to phenylalanine (Y179F) TAC → TTC; and serine-180 to alanine (S180A) TCA → GCA. Amino acid numbering is according to human PGRMC1 Uniprot O00264, which does not include the initiator methionine. Uniprot Q6IB11 corresponds to the same sequence with initiator methionine included, in which mutated human PGRMC1 amino acids would be numbered as S57, C129, Y139, Y180, and S181. The position of three tandem influenza virus hemaglutinin epitope sequences (HA) is indicated at the carboxyl-terminus of the open reading frame. PGMRC, progesterone receptor membrane component.