Proteomic Analysis of Ferrochelatase Interactome in Erythroid and Non-Erythroid Cells
Heme is an essential cofactor for multiple cellular processes in most organisms. In developing erythroid cells, the demand for heme synthesis is high, but is significantly lower in non-erythroid cells. While the biosynthesis of heme in metazoans is well understood, the tissue-specific regulation of...
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MDPI AG
2023-02-01
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| Series: | Life |
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| Online Access: | https://www.mdpi.com/2075-1729/13/2/577 |
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| author | Chibuike David Obi Harry A. Dailey Yasaman Jami-Alahmadi James A. Wohlschlegel Amy E. Medlock |
| author_facet | Chibuike David Obi Harry A. Dailey Yasaman Jami-Alahmadi James A. Wohlschlegel Amy E. Medlock |
| author_sort | Chibuike David Obi |
| collection | DOAJ |
| description | Heme is an essential cofactor for multiple cellular processes in most organisms. In developing erythroid cells, the demand for heme synthesis is high, but is significantly lower in non-erythroid cells. While the biosynthesis of heme in metazoans is well understood, the tissue-specific regulation of the pathway is less explored. To better understand this, we analyzed the mitochondrial heme metabolon in erythroid and non-erythroid cell lines from the perspective of ferrochelatase (FECH), the terminal enzyme in the heme biosynthetic pathway. Affinity purification of FLAG-tagged-FECH, together with mass spectrometric analysis, was carried out to identify putative protein partners in human and murine cell lines. Proteins involved in the heme biosynthetic process and mitochondrial organization were identified as the core components of the FECH interactome. Interestingly, in non-erythroid cell lines, the FECH interactome is highly enriched with proteins associated with the tricarboxylic acid (TCA) cycle. Overall, our study shows that the mitochondrial heme metabolon in erythroid and non-erythroid cells has similarities and differences, and suggests new roles for the mitochondrial heme metabolon and heme in regulating metabolic flux and key cellular processes. |
| first_indexed | 2024-03-11T08:33:08Z |
| format | Article |
| id | doaj.art-fef55fd4c8ec4d3392a2e520ba35cd14 |
| institution | Directory Open Access Journal |
| issn | 2075-1729 |
| language | English |
| last_indexed | 2024-03-11T08:33:08Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Life |
| spelling | doaj.art-fef55fd4c8ec4d3392a2e520ba35cd142023-11-16T21:43:11ZengMDPI AGLife2075-17292023-02-0113257710.3390/life13020577Proteomic Analysis of Ferrochelatase Interactome in Erythroid and Non-Erythroid CellsChibuike David Obi0Harry A. Dailey1Yasaman Jami-Alahmadi2James A. Wohlschlegel3Amy E. Medlock4Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USADepartment of Biological Chemistry, University of California Los Angeles, Los Angeles, CA 90095, USADepartment of Biological Chemistry, University of California Los Angeles, Los Angeles, CA 90095, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USAHeme is an essential cofactor for multiple cellular processes in most organisms. In developing erythroid cells, the demand for heme synthesis is high, but is significantly lower in non-erythroid cells. While the biosynthesis of heme in metazoans is well understood, the tissue-specific regulation of the pathway is less explored. To better understand this, we analyzed the mitochondrial heme metabolon in erythroid and non-erythroid cell lines from the perspective of ferrochelatase (FECH), the terminal enzyme in the heme biosynthetic pathway. Affinity purification of FLAG-tagged-FECH, together with mass spectrometric analysis, was carried out to identify putative protein partners in human and murine cell lines. Proteins involved in the heme biosynthetic process and mitochondrial organization were identified as the core components of the FECH interactome. Interestingly, in non-erythroid cell lines, the FECH interactome is highly enriched with proteins associated with the tricarboxylic acid (TCA) cycle. Overall, our study shows that the mitochondrial heme metabolon in erythroid and non-erythroid cells has similarities and differences, and suggests new roles for the mitochondrial heme metabolon and heme in regulating metabolic flux and key cellular processes.https://www.mdpi.com/2075-1729/13/2/577erythroidferrochelatasehemeheme biosynthesisinteractomemetabolon |
| spellingShingle | Chibuike David Obi Harry A. Dailey Yasaman Jami-Alahmadi James A. Wohlschlegel Amy E. Medlock Proteomic Analysis of Ferrochelatase Interactome in Erythroid and Non-Erythroid Cells Life erythroid ferrochelatase heme heme biosynthesis interactome metabolon |
| title | Proteomic Analysis of Ferrochelatase Interactome in Erythroid and Non-Erythroid Cells |
| title_full | Proteomic Analysis of Ferrochelatase Interactome in Erythroid and Non-Erythroid Cells |
| title_fullStr | Proteomic Analysis of Ferrochelatase Interactome in Erythroid and Non-Erythroid Cells |
| title_full_unstemmed | Proteomic Analysis of Ferrochelatase Interactome in Erythroid and Non-Erythroid Cells |
| title_short | Proteomic Analysis of Ferrochelatase Interactome in Erythroid and Non-Erythroid Cells |
| title_sort | proteomic analysis of ferrochelatase interactome in erythroid and non erythroid cells |
| topic | erythroid ferrochelatase heme heme biosynthesis interactome metabolon |
| url | https://www.mdpi.com/2075-1729/13/2/577 |
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