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Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids

Polyneuropathy, hearing loss, ataxia, retinitis pigmentosa, and cataract (PHARC) is a rare genetic human neurological disorder caused by null mutations to the Abhd12 gene, which encodes the integral membrane serine hydrolase enzyme ABHD12. Although the role that ABHD12 plays in PHARC is understood,...

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Autores principales: Joshi, Alaumy, Shaikh, Minhaj, Singh, Shubham, Rajendran, Abinaya, Mhetre, Amol, Kamat, Siddhesh S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6217928/
https://www.ncbi.nlm.nih.gov/pubmed/30237167
http://dx.doi.org/10.1074/jbc.RA118.005640
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author Joshi, Alaumy
Shaikh, Minhaj
Singh, Shubham
Rajendran, Abinaya
Mhetre, Amol
Kamat, Siddhesh S.
author_facet Joshi, Alaumy
Shaikh, Minhaj
Singh, Shubham
Rajendran, Abinaya
Mhetre, Amol
Kamat, Siddhesh S.
author_sort Joshi, Alaumy
collection PubMed
description Polyneuropathy, hearing loss, ataxia, retinitis pigmentosa, and cataract (PHARC) is a rare genetic human neurological disorder caused by null mutations to the Abhd12 gene, which encodes the integral membrane serine hydrolase enzyme ABHD12. Although the role that ABHD12 plays in PHARC is understood, the thorough biochemical characterization of ABHD12 is lacking. Here, we report the facile synthesis of mono-1-(fatty)acyl-glycerol lipids of varying chain lengths and unsaturation and use this lipid substrate library to biochemically characterize recombinant mammalian ABHD12. The substrate profiling study for ABHD12 suggested that this enzyme requires glycosylation for optimal activity and that it has a strong preference for very-long-chain lipid substrates. We further validated this substrate profile against brain membrane lysates generated from WT and ABHD12 knockout mice. Finally, using cellular organelle fractionation and immunofluorescence assays, we show that mammalian ABHD12 is enriched on the endoplasmic reticulum membrane, where most of the very-long-chain fatty acids are biosynthesized in cells. Taken together, our findings provide a biochemical explanation for why very-long-chain lipids (such as lysophosphatidylserine lipids) accumulate in the brains of ABHD12 knockout mice, which is a murine model of PHARC.
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spelling pubmed-62179282018-11-05 Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids Joshi, Alaumy Shaikh, Minhaj Singh, Shubham Rajendran, Abinaya Mhetre, Amol Kamat, Siddhesh S. J Biol Chem Lipids Polyneuropathy, hearing loss, ataxia, retinitis pigmentosa, and cataract (PHARC) is a rare genetic human neurological disorder caused by null mutations to the Abhd12 gene, which encodes the integral membrane serine hydrolase enzyme ABHD12. Although the role that ABHD12 plays in PHARC is understood, the thorough biochemical characterization of ABHD12 is lacking. Here, we report the facile synthesis of mono-1-(fatty)acyl-glycerol lipids of varying chain lengths and unsaturation and use this lipid substrate library to biochemically characterize recombinant mammalian ABHD12. The substrate profiling study for ABHD12 suggested that this enzyme requires glycosylation for optimal activity and that it has a strong preference for very-long-chain lipid substrates. We further validated this substrate profile against brain membrane lysates generated from WT and ABHD12 knockout mice. Finally, using cellular organelle fractionation and immunofluorescence assays, we show that mammalian ABHD12 is enriched on the endoplasmic reticulum membrane, where most of the very-long-chain fatty acids are biosynthesized in cells. Taken together, our findings provide a biochemical explanation for why very-long-chain lipids (such as lysophosphatidylserine lipids) accumulate in the brains of ABHD12 knockout mice, which is a murine model of PHARC. American Society for Biochemistry and Molecular Biology 2018-11-02 2018-09-20 /pmc/articles/PMC6217928/ /pubmed/30237167 http://dx.doi.org/10.1074/jbc.RA118.005640 Text en © 2018 Joshi et al. Author's Choice—Final version open access under the terms of the Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0) .
spellingShingle Lipids
Joshi, Alaumy
Shaikh, Minhaj
Singh, Shubham
Rajendran, Abinaya
Mhetre, Amol
Kamat, Siddhesh S.
Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids
title Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids
title_full Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids
title_fullStr Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids
title_full_unstemmed Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids
title_short Biochemical characterization of the PHARC-associated serine hydrolase ABHD12 reveals its preference for very-long-chain lipids
title_sort biochemical characterization of the pharc-associated serine hydrolase abhd12 reveals its preference for very-long-chain lipids
topic Lipids
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6217928/
https://www.ncbi.nlm.nih.gov/pubmed/30237167
http://dx.doi.org/10.1074/jbc.RA118.005640
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