Cardiolipin Stabilizes and Increases Catalytic Efficiency of Carnitine Palmitoyltransferase II and Its Variants S113L, P50H, and Y479F
Muscle carnitine palmitoyltransferase II (CPT II) deficiency is associated with various mutations in <i>CPT2</i> gene. In the present study, the impact of the two CPT II variants P50H and Y479F were characterized in terms of stability and activity in vitro in comparison to wildtype (WT)...
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MDPI AG
2021-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/9/4831 |
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| author | Beate Meinhardt Leila Motlagh Scholle Franziska Seifert Martina Anwand Markus Pietzsch Stephan Zierz |
| author_facet | Beate Meinhardt Leila Motlagh Scholle Franziska Seifert Martina Anwand Markus Pietzsch Stephan Zierz |
| author_sort | Beate Meinhardt |
| collection | DOAJ |
| description | Muscle carnitine palmitoyltransferase II (CPT II) deficiency is associated with various mutations in <i>CPT2</i> gene. In the present study, the impact of the two CPT II variants P50H and Y479F were characterized in terms of stability and activity in vitro in comparison to wildtype (WT) and the well investigated variant S113L. While the initial enzyme activity of all variants showed wild-type-like behavior, the activity half-lives of the variants at different temperatures were severely reduced. This finding was validated by the investigation of thermostability of the enzymes using nano differential scanning fluorimetry (nanoDSF). Further, it was studied whether the protein stabilizing diphosphatidylglycerol cardiolipin (CL) has an effect on the variants. CL indeed had a positive effect on the stability. This effect was strongest for WT and least pronounced for variant P50H. Additionally, CL improved the catalytic efficiency for CPT II WT and the investigated variants by twofold when carnitine was the varied substrate due to a decrease in K<sub>M</sub>. However, there was no influence detected for the variation of substrate palmitoyl-CoA. The functional consequences of the stabilization by CL in vivo remain open. |
| first_indexed | 2024-03-10T11:44:42Z |
| format | Article |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T11:44:42Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-da9ef6ced4094f49a581428d7ee918852023-11-21T18:11:54ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-05-01229483110.3390/ijms22094831Cardiolipin Stabilizes and Increases Catalytic Efficiency of Carnitine Palmitoyltransferase II and Its Variants S113L, P50H, and Y479FBeate Meinhardt0Leila Motlagh Scholle1Franziska Seifert2Martina Anwand3Markus Pietzsch4Stephan Zierz5Department of Neurology, Martin-Luther-University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), GermanyDepartment of Neurology, Martin-Luther-University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), GermanyDepartment of Pharmaceutical Technology and Biopharmacy, Institute of Pharmacy, Martin-Luther-University Halle-Wittenberg, Weinbergweg 22, 06120 Halle (Saale), GermanyDepartment of Pharmaceutical Technology and Biopharmacy, Institute of Pharmacy, Martin-Luther-University Halle-Wittenberg, Weinbergweg 22, 06120 Halle (Saale), GermanyDepartment of Pharmaceutical Technology and Biopharmacy, Institute of Pharmacy, Martin-Luther-University Halle-Wittenberg, Weinbergweg 22, 06120 Halle (Saale), GermanyDepartment of Neurology, Martin-Luther-University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), GermanyMuscle carnitine palmitoyltransferase II (CPT II) deficiency is associated with various mutations in <i>CPT2</i> gene. In the present study, the impact of the two CPT II variants P50H and Y479F were characterized in terms of stability and activity in vitro in comparison to wildtype (WT) and the well investigated variant S113L. While the initial enzyme activity of all variants showed wild-type-like behavior, the activity half-lives of the variants at different temperatures were severely reduced. This finding was validated by the investigation of thermostability of the enzymes using nano differential scanning fluorimetry (nanoDSF). Further, it was studied whether the protein stabilizing diphosphatidylglycerol cardiolipin (CL) has an effect on the variants. CL indeed had a positive effect on the stability. This effect was strongest for WT and least pronounced for variant P50H. Additionally, CL improved the catalytic efficiency for CPT II WT and the investigated variants by twofold when carnitine was the varied substrate due to a decrease in K<sub>M</sub>. However, there was no influence detected for the variation of substrate palmitoyl-CoA. The functional consequences of the stabilization by CL in vivo remain open.https://www.mdpi.com/1422-0067/22/9/4831carnitine palmitoyltransferase IIcardiolipinthermostabilityS113LP50HY479F |
| spellingShingle | Beate Meinhardt Leila Motlagh Scholle Franziska Seifert Martina Anwand Markus Pietzsch Stephan Zierz Cardiolipin Stabilizes and Increases Catalytic Efficiency of Carnitine Palmitoyltransferase II and Its Variants S113L, P50H, and Y479F International Journal of Molecular Sciences carnitine palmitoyltransferase II cardiolipin thermostability S113L P50H Y479F |
| title | Cardiolipin Stabilizes and Increases Catalytic Efficiency of Carnitine Palmitoyltransferase II and Its Variants S113L, P50H, and Y479F |
| title_full | Cardiolipin Stabilizes and Increases Catalytic Efficiency of Carnitine Palmitoyltransferase II and Its Variants S113L, P50H, and Y479F |
| title_fullStr | Cardiolipin Stabilizes and Increases Catalytic Efficiency of Carnitine Palmitoyltransferase II and Its Variants S113L, P50H, and Y479F |
| title_full_unstemmed | Cardiolipin Stabilizes and Increases Catalytic Efficiency of Carnitine Palmitoyltransferase II and Its Variants S113L, P50H, and Y479F |
| title_short | Cardiolipin Stabilizes and Increases Catalytic Efficiency of Carnitine Palmitoyltransferase II and Its Variants S113L, P50H, and Y479F |
| title_sort | cardiolipin stabilizes and increases catalytic efficiency of carnitine palmitoyltransferase ii and its variants s113l p50h and y479f |
| topic | carnitine palmitoyltransferase II cardiolipin thermostability S113L P50H Y479F |
| url | https://www.mdpi.com/1422-0067/22/9/4831 |
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