Sigma-2 receptor and progesterone receptor membrane component 1 (PGRMC1) are two different proteins: Proofs by fluorescent labeling and binding of sigma-2 receptor ligands to PGRMC1

Maria Laura Pati, Diana Groza, Chiara Riganti, Joanna Kopecka, Mauro Niso, Francesco Berardi, Sonja Hager, Petra Heffeter, Miwa Hirai, Hitoshi Tsugawa, Yasuaki Kabe, Makoto Suematsu, Carmen Abate

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

A controversial relationship between sigma-2 and progesterone receptor membrane component 1 (PGRMC1) proteins, both representing promising targets for the therapy and diagnosis of tumors, exists since 2011, when the sigma-2 receptor was reported to be identical to PGRMC1. Because a misidentification of these proteins will lead to biased future research hampering the possible diagnostic and therapeutic exploitation of the two targets, there is the need to solve the debate on their identity. With this aim, we have herein investigated uptake and distribution of structurally different fluorescent sigma-2 receptor ligands by flow cytometry and confocal microscopy in MCF7 cells, where together with intrinsic sigma-2 receptors, PGRMC1 was constitutively present or alternatively silenced or overexpressed. HCT116 cells, with constitutive or silenced PGRMC1, were also studied. These experiments showed that the fluorescent sigma-2 ligands bind to their receptor irrespective of PGRMC1 expression. Furthermore, isothermal titration calorimetry was conducted to examine if DTG and PB28, two structurally distinct nanomolar affinity sigma-2 ligands, bind to purified PGRMC1 proteins that have recently been revealed to form both apo-monomeric and heme-mediated dimeric forms. While no binding to apo-PGRMC1 monomer was detected, a micromolar affinity to heme-mediated dimerized PGRMC1 was demonstrated in DTG but not in PB28. The current data provide evidence that sigma-2 receptor and PGRMC1 are not identical, paving the pathway for future unbiased research in which these two attractive targets are treated as different proteins while the identification of the true sigma-2 protein further needs to be pursued.

Original languageEnglish
Pages (from-to)67-74
Number of pages8
JournalPharmacological Research
Volume117
DOIs
Publication statusPublished - 2017 Mar 1

Fingerprint

Progesterone Receptors
Ligands
Membranes
Proteins
Heme
HCT116 Cells
sigma-2 receptor
Calorimetry
MCF-7 Cells
Confocal Microscopy
Flow Cytometry
Therapeutics
Research

Keywords

  • apo-PGRMC1 monomer
  • Confocal microscopy
  • Flow-cytometry
  • Heme-PGRMC1 dimer
  • PGRMC1
  • Sigma-2 receptor

ASJC Scopus subject areas

  • Pharmacology

Cite this

Sigma-2 receptor and progesterone receptor membrane component 1 (PGRMC1) are two different proteins : Proofs by fluorescent labeling and binding of sigma-2 receptor ligands to PGRMC1. / Pati, Maria Laura; Groza, Diana; Riganti, Chiara; Kopecka, Joanna; Niso, Mauro; Berardi, Francesco; Hager, Sonja; Heffeter, Petra; Hirai, Miwa; Tsugawa, Hitoshi; Kabe, Yasuaki; Suematsu, Makoto; Abate, Carmen.

In: Pharmacological Research, Vol. 117, 01.03.2017, p. 67-74.

Research output: Contribution to journalArticle

Pati, Maria Laura ; Groza, Diana ; Riganti, Chiara ; Kopecka, Joanna ; Niso, Mauro ; Berardi, Francesco ; Hager, Sonja ; Heffeter, Petra ; Hirai, Miwa ; Tsugawa, Hitoshi ; Kabe, Yasuaki ; Suematsu, Makoto ; Abate, Carmen. / Sigma-2 receptor and progesterone receptor membrane component 1 (PGRMC1) are two different proteins : Proofs by fluorescent labeling and binding of sigma-2 receptor ligands to PGRMC1. In: Pharmacological Research. 2017 ; Vol. 117. pp. 67-74.
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AU - Niso, Mauro

AU - Berardi, Francesco

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AU - Heffeter, Petra

AU - Hirai, Miwa

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AU - Kabe, Yasuaki

AU - Suematsu, Makoto

AU - Abate, Carmen

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