Defining Requirements for Heme Binding in PGRMC1 and Identifying Key Elements that Influence Protein Dimerization

Abstract

Progesterone receptor membrane component 1 (PGRMC1) binds heme via a surface-exposed site and displays some structural resemblance to cytochrome b5 despite their different functions. In the case of PGRMC1, it is the protein interaction with drug-metabolizing cytochrome P450s and the epidermal growth factor receptor that has garnered the most attention. These interactions are thought to result in a compromised ability to metabolize common chemotherapy agents and to enhance cancer cell proliferation. X-ray crystallography and immunoprecipitation data have suggested that heme-mediated PGRMC1 dimers are important for facilitating these interactions. However, more recent studies have called into question the requirement of heme binding for PGRMC1 dimerization. Our study employs spectroscopic and computational methods to probe and define heme binding and its impact on PGRMC1 dimerization. Fluorescence, electron paramagnetic resonance and circular dichroism spectroscopies confirm heme binding to apo-PGRMC1 and were used to demonstrate the stabilizing effect of heme on the wild-type protein. We also utilized variants (C129S and Y113F) to precisely define the contributions of disulfide bonds and direct heme coordination to PGRMC1 dimerization. Understanding the key factors involved in these processes has important implications for downstream protein−protein interactions that may influence the metabolism of chemotherapeutic agents. This work opens avenues for deeper exploration into the physiological significance of the truncated-PGRMC1 model and developing design principles for potential therapeutics to target PGRMC1 dimerization and downstream interactions.

Pleiotropy of Progesterone Receptor Membrane Component 1 in Modulation of Cytochrome P450 Activity

“…Moreover, although a disulfide bond between the two Cys129 residues had been observed in the crystal of PGRMC1 [ 151 ], the mutants previously assessed (Cys129Ala and Cys129Ser) had shown no significantly diminished heme absorption and dimerization [ 130 , 151 ]. However, as highlighted by new structural insights, the impact of the disulfide bond on PGRMC1 dimerization in the absence of heme had not been investigated [ 261 ]. The recent study proposes intermolecular disulfide bond formation through the previously overlooked Cys129 as another route for PGRMC1 dimer formation.…”