The Sigma-2 Receptor and Progesterone Receptor Membrane Component 1 are Different Binding Sites Derived From Independent Genes

摘要

The sigma-2 receptor (S2R) is a potential therapeutic target for cancer and neuronal diseases. However, the identity of the S2R has remained a matter of debate. Historically, the S2R has been defined as (1) a binding site with high affinity to 1,3-di-o-tolylguanidine (DTG) and haloperidol but not to the selective sigma-1 receptor ligand (+)-pentazocine, and (2) a protein of 18-21kDa, as shown by specific photolabeling with [^3H]-Azido-DTG and [^1^2^5I]-iodoazido-fenpropimorph ([^1^2^5I]-IAF). Recently, the progesterone receptor membrane component 1 (PGRMC1), a 25kDa protein, was reported to be the S2R (Nature Communications, 2011, 2:380). To confirm this identification, we created PGRMC1 knockout NSC34 cell lines using the CRISPR/Cas9 technology. We found that in NSC34 cells devoid of or overexpressing PGRMC1, the maximum [^3H]-DTG binding to the S2R (B"m"a"x) as well as the DTG-protectable [^1^2^5I]-IAF photolabeling of the S2R were similar to those of wild-type control cells. Furthermore, the affinities of DTG and haloperidol for PGRMC1 (K"I=472@mM and 350@mM, respectively), as determined in competition with [^3H]-progesterone, were more than 3 orders of magnitude lower than those reported for the S2R (20-80nM). These results clarify that PGRMC1 and the S2R are distinct binding sites expressed by different genes.