A Novel Role for Progesterone Receptor Membrane Component 1 (PGRMC1): A Partner and Regulator of Ferrochelatase

Robert B. Piel, Mesafint T. Shiferaw, Ajay A. Vashisht, Jason R. Marcero, Jeremy L. Praissman, John D. Phillips, James A. Wohlschlegel, Amy E. Medlock

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

Heme is an iron-containing cofactor essential for multiple cellular processes and fundamental activities such as oxygen transport. To better understand the means by which heme synthesis is regulated during erythropoiesis, affinity purification coupled with mass spectrometry (MS) was performed to identify putative protein partners interacting with ferrochelatase (FECH), the terminal enzyme in the heme biosynthetic pathway. Both progesterone receptor membrane component 1 (PGRMC1) and progesterone receptor membrane component 2 (PGRMC2) were identified in these experiments. These interactions were validated by reciprocal affinity purification followed by MS analysis and immunoblotting. The interaction between PGRMC1 and FECH was confirmed in vitro and in HEK 293T cells, a non-erythroid cell line. When cells that are recognized models for erythroid differentiation were treated with a small molecule inhibitor of PGRMC1, AG-205, there was an observed decrease in the level of hemoglobinization relative to that of untreated cells. In vitro heme transfer experiments showed that purified PGRMC1 was able to donate heme to apo-cytochrome b5. In the presence of PGRMC1, in vitro measured FECH activity decreased in a dose-dependent manner. Interactions between FECH and PGRMC1 were strongest for the conformation of FECH associated with product release, suggesting that PGRMC1 may regulate FECH activity by controlling heme release. Overall, the data illustrate a role for PGRMC1 in regulating heme synthesis via interactions with FECH and suggest that PGRMC1 may be a heme chaperone or sensor.

Original languageEnglish (US)
Pages (from-to)5204-5217
Number of pages14
JournalBiochemistry
Volume55
Issue number37
DOIs
StatePublished - Sep 20 2016

ASJC Scopus subject areas

  • Biochemistry

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    Piel, R. B., Shiferaw, M. T., Vashisht, A. A., Marcero, J. R., Praissman, J. L., Phillips, J. D., Wohlschlegel, J. A., & Medlock, A. E. (2016). A Novel Role for Progesterone Receptor Membrane Component 1 (PGRMC1): A Partner and Regulator of Ferrochelatase. Biochemistry, 55(37), 5204-5217. https://doi.org/10.1021/acs.biochem.6b00756