Proteomics

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LC-MSMS with isobaric tagging of Pgrmc1 KO mouse liver membranes expressing Flag-Pgrmc1 or Y113F Flag-Pgrmc1

ABSTRACT: Progesterone receptor membrane component 1 (PGRMC1) is a heme binding protein implicated in a wide range of cellular functions. Our previous studies showed that PGRMC1 binds to cytochromes P450 in yeast and mammalian cells and promotes activity of these enzymes. Recently, the paralog PGRMC2 was shown to function as an intracellular heme chaperone. Here, we examined the function of the Pgrmc1 Y113F mutant that is defective for heme iron coordination by measuring levels of membrane proteins in livers from Pgrmc1 knockout mice infected with AAV8 expressing either GFP, Flag-Pgrmc1, or Y113F Flag-Pgrmc1. These experiments (1) confirmed findings in uninfected wild-type and Pgrmc1 knockout mice and (2) revealed that Pgrmc1 Y113F functions to stabilize cytochrome P450 enzymes similar to wild-type Pgrmc1.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Liver

SUBMITTER: Peter Espenshade  

LAB HEAD: Peter J. Espenshade

PROVIDER: PXD028288 | Pride | 2022-02-16

REPOSITORIES: Pride

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Progesterone receptor membrane component 1 (PGRMC1) binds and stabilizes cytochromes P450 through a heme-independent mechanism.

McGuire Meredith R MR   Mukhopadhyay Debaditya D   Myers Stephanie L SL   Mosher Eric P EP   Brookheart Rita T RT   Kammers Kai K   Sehgal Alfica A   Selen Ebru S ES   Wolfgang Michael J MJ   Bumpus Namandjé N NN   Espenshade Peter J PJ  

The Journal of biological chemistry 20211020 5

Progesterone receptor membrane component 1 (PGRMC1) is a heme-binding protein implicated in a wide range of cellular functions. We previously showed that PGRMC1 binds to cytochromes P450 in yeast and mammalian cells and supports their activity. Recently, the paralog PGRMC2 was shown to function as a heme chaperone. The extent of PGRMC1 function in cytochrome P450 biology and whether PGRMC1 is also a heme chaperone are unknown. Here, we examined the function of Pgrmc1 in mouse liver using a knock  ...[more]

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