The Multienzyme Complex Nature of Dehydroepiandrosterone Sulfate Biosynthesis
Dehydroepiandrosterone (DHEA), a precursor of steroid sex hormones, is synthesized by steroid 17-alpha-hydroxylase/17,20-lyase (CYP17A1) with the participation of microsomal cytochrome b5 (CYB5A) and cytochrome P450 reductase (CPR), followed by sulfation by two cytosolic sulfotransferases, SULT1E1 a...
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| Format: | Article |
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MDPI AG
2024-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/4/2072 |
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| author | Anastasiya Tumilovich Evgeniy Yablokov Yuri Mezentsev Pavel Ershov Viktoriia Basina Oksana Gnedenko Leonid Kaluzhskiy Tatsiana Tsybruk Irina Grabovec Maryia Kisel Polina Shabunya Natalia Soloveva Nikita Vavilov Andrei Gilep Alexis Ivanov |
| author_facet | Anastasiya Tumilovich Evgeniy Yablokov Yuri Mezentsev Pavel Ershov Viktoriia Basina Oksana Gnedenko Leonid Kaluzhskiy Tatsiana Tsybruk Irina Grabovec Maryia Kisel Polina Shabunya Natalia Soloveva Nikita Vavilov Andrei Gilep Alexis Ivanov |
| author_sort | Anastasiya Tumilovich |
| collection | DOAJ |
| description | Dehydroepiandrosterone (DHEA), a precursor of steroid sex hormones, is synthesized by steroid 17-alpha-hydroxylase/17,20-lyase (CYP17A1) with the participation of microsomal cytochrome b5 (CYB5A) and cytochrome P450 reductase (CPR), followed by sulfation by two cytosolic sulfotransferases, SULT1E1 and SULT2A1, for storage and transport to tissues in which its synthesis is not available. The involvement of CYP17A1 and SULTs in these successive reactions led us to consider the possible interaction of SULTs with DHEA-producing CYP17A1 and its redox partners. Text mining analysis, protein–protein network analysis, and gene co-expression analysis were performed to determine the relationships between SULTs and microsomal CYP isoforms. For the first time, using surface plasmon resonance, we detected interactions between CYP17A1 and SULT2A1 or SULT1E1. SULTs also interacted with CYB5A and CPR. The interaction parameters of SULT2A1/CYP17A1 and SULT2A1/CYB5A complexes seemed to be modulated by 3′-phosphoadenosine-5′-phosphosulfate (PAPS). Affinity purification, combined with mass spectrometry (AP-MS), allowed us to identify a spectrum of SULT1E1 potential protein partners, including CYB5A. We showed that the enzymatic activity of SULTs increased in the presence of only CYP17A1 or CYP17A1 and CYB5A mixture. The structures of CYP17A1/SULT1E1 and CYB5A/SULT1E1 complexes were predicted. Our data provide novel fundamental information about the organization of microsomal CYP-dependent macromolecular complexes. |
| first_indexed | 2024-03-07T22:30:04Z |
| format | Article |
| id | doaj.art-d17b52e100b34eb98ae64a4985b5e9f4 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-07T22:30:04Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d17b52e100b34eb98ae64a4985b5e9f42024-02-23T15:19:38ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-02-01254207210.3390/ijms25042072The Multienzyme Complex Nature of Dehydroepiandrosterone Sulfate BiosynthesisAnastasiya Tumilovich0Evgeniy Yablokov1Yuri Mezentsev2Pavel Ershov3Viktoriia Basina4Oksana Gnedenko5Leonid Kaluzhskiy6Tatsiana Tsybruk7Irina Grabovec8Maryia Kisel9Polina Shabunya10Natalia Soloveva11Nikita Vavilov12Andrei Gilep13Alexis Ivanov14Institute of Bioorganic Chemistry NASB, 5 Building 2, V.F. Kuprevich Street, 220141 Minsk, BelarusInstitute of Biomedical Chemistry, 10 Building 8, Pogodinskaya Street, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, 10 Building 8, Pogodinskaya Street, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, 10 Building 8, Pogodinskaya Street, 119121 Moscow, RussiaResearch Centre for Medical Genetics, 1 Moskvorechye Street, 115522 Moscow, RussiaInstitute of Biomedical Chemistry, 10 Building 8, Pogodinskaya Street, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, 10 Building 8, Pogodinskaya Street, 119121 Moscow, RussiaInstitute of Bioorganic Chemistry NASB, 5 Building 2, V.F. Kuprevich Street, 220141 Minsk, BelarusInstitute of Bioorganic Chemistry NASB, 5 Building 2, V.F. Kuprevich Street, 220141 Minsk, BelarusInstitute of Bioorganic Chemistry NASB, 5 Building 2, V.F. Kuprevich Street, 220141 Minsk, BelarusInstitute of Bioorganic Chemistry NASB, 5 Building 2, V.F. Kuprevich Street, 220141 Minsk, BelarusInstitute of Biomedical Chemistry, 10 Building 8, Pogodinskaya Street, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, 10 Building 8, Pogodinskaya Street, 119121 Moscow, RussiaInstitute of Bioorganic Chemistry NASB, 5 Building 2, V.F. Kuprevich Street, 220141 Minsk, BelarusInstitute of Biomedical Chemistry, 10 Building 8, Pogodinskaya Street, 119121 Moscow, RussiaDehydroepiandrosterone (DHEA), a precursor of steroid sex hormones, is synthesized by steroid 17-alpha-hydroxylase/17,20-lyase (CYP17A1) with the participation of microsomal cytochrome b5 (CYB5A) and cytochrome P450 reductase (CPR), followed by sulfation by two cytosolic sulfotransferases, SULT1E1 and SULT2A1, for storage and transport to tissues in which its synthesis is not available. The involvement of CYP17A1 and SULTs in these successive reactions led us to consider the possible interaction of SULTs with DHEA-producing CYP17A1 and its redox partners. Text mining analysis, protein–protein network analysis, and gene co-expression analysis were performed to determine the relationships between SULTs and microsomal CYP isoforms. For the first time, using surface plasmon resonance, we detected interactions between CYP17A1 and SULT2A1 or SULT1E1. SULTs also interacted with CYB5A and CPR. The interaction parameters of SULT2A1/CYP17A1 and SULT2A1/CYB5A complexes seemed to be modulated by 3′-phosphoadenosine-5′-phosphosulfate (PAPS). Affinity purification, combined with mass spectrometry (AP-MS), allowed us to identify a spectrum of SULT1E1 potential protein partners, including CYB5A. We showed that the enzymatic activity of SULTs increased in the presence of only CYP17A1 or CYP17A1 and CYB5A mixture. The structures of CYP17A1/SULT1E1 and CYB5A/SULT1E1 complexes were predicted. Our data provide novel fundamental information about the organization of microsomal CYP-dependent macromolecular complexes.https://www.mdpi.com/1422-0067/25/4/2072cytochrome P450cytosolic sulfotransferasecytochrome b5surface plasmon resonanceenzymatic assayin silico modeling |
| spellingShingle | Anastasiya Tumilovich Evgeniy Yablokov Yuri Mezentsev Pavel Ershov Viktoriia Basina Oksana Gnedenko Leonid Kaluzhskiy Tatsiana Tsybruk Irina Grabovec Maryia Kisel Polina Shabunya Natalia Soloveva Nikita Vavilov Andrei Gilep Alexis Ivanov The Multienzyme Complex Nature of Dehydroepiandrosterone Sulfate Biosynthesis International Journal of Molecular Sciences cytochrome P450 cytosolic sulfotransferase cytochrome b5 surface plasmon resonance enzymatic assay in silico modeling |
| title | The Multienzyme Complex Nature of Dehydroepiandrosterone Sulfate Biosynthesis |
| title_full | The Multienzyme Complex Nature of Dehydroepiandrosterone Sulfate Biosynthesis |
| title_fullStr | The Multienzyme Complex Nature of Dehydroepiandrosterone Sulfate Biosynthesis |
| title_full_unstemmed | The Multienzyme Complex Nature of Dehydroepiandrosterone Sulfate Biosynthesis |
| title_short | The Multienzyme Complex Nature of Dehydroepiandrosterone Sulfate Biosynthesis |
| title_sort | multienzyme complex nature of dehydroepiandrosterone sulfate biosynthesis |
| topic | cytochrome P450 cytosolic sulfotransferase cytochrome b5 surface plasmon resonance enzymatic assay in silico modeling |
| url | https://www.mdpi.com/1422-0067/25/4/2072 |
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