Biochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).

<h4>Background</h4>Human lymphocyte antigen B-associated transcript 5 (BAT5, also known as ABHD16A) is a poorly characterized 63 kDa protein belonging to the α/β-hydrolase domain (ABHD) containing family of metabolic serine hydrolases. Its natural substrates and biochemical properties ar...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Juha R Savinainen, Jayendra Z Patel, Teija Parkkari, Dina Navia-Paldanius, Joona J T Marjamaa, Tuomo Laitinen, Tapio Nevalainen, Jarmo T Laitinen
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7badfbd1be224b0b86ab6594956a58a2
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:7badfbd1be224b0b86ab6594956a58a2
record_format	dspace
spelling	oai:doaj.org-article:7badfbd1be224b0b86ab6594956a58a22021-11-25T05:57:38ZBiochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).1932-620310.1371/journal.pone.0109869https://doaj.org/article/7badfbd1be224b0b86ab6594956a58a22014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0109869https://doaj.org/toc/1932-6203<h4>Background</h4>Human lymphocyte antigen B-associated transcript 5 (BAT5, also known as ABHD16A) is a poorly characterized 63 kDa protein belonging to the α/β-hydrolase domain (ABHD) containing family of metabolic serine hydrolases. Its natural substrates and biochemical properties are unknown.<h4>Methodology/principal findings</h4>Amino acid sequence comparison between seven mammalian BAT5 orthologs revealed that the overall primary structure was highly (≥95%) conserved. Activity-based protein profiling (ABPP) confirmed successful generation of catalytically active human (h) and mouse (m) BAT5 in HEK293 cells, enabling further biochemical characterization. A sensitive fluorescent glycerol assay reported hBAT5-mediated hydrolysis of medium-chain saturated (C14:0), long-chain unsaturated (C18:1, C18:2, C20:4) monoacylglycerols (MAGs) and 15-deoxy-Δ12,14-prostaglandin J2-2-glycerol ester (15d-PGJ2-G). In contrast, hBAT5 possessed only marginal diacylglycerol (DAG), triacylglycerol (TAG), or lysophospholipase activity. The best MAG substrates were 1-linoleylglycerol (1-LG) and 15d-PGJ2-G, both exhibiting low-micromolar Km values. BAT5 had a neutral pH optimum and showed preference for the 1(3)- vs. 2-isomers of MAGs C18:1, C18:2 and C20:4. Inhibitor profiling revealed that β-lactone-based lipase inhibitors were nanomolar inhibitors of hBAT5 activity (palmostatin B > tetrahydrolipstatin > ebelactone A). Moreover, the hormone-sensitive lipase inhibitor C7600 (5-methoxy-3-(4-phenoxyphenyl)-3H-[1], [3], [4]oxadiazol-2-one) was identified as a highly potent inhibitor (IC50 8.3 nM). Phenyl and benzyl substituted analogs of C7600 with increased BAT5 selectivity were synthesized and a preliminary SAR analysis was conducted to obtain initial insights into the active site dimensions.<h4>Conclusions/significance</h4>This study provides an initial characterization of BAT5 activity, unveiling the biochemical and pharmacological properties with in vitro substrate preferences and inhibitor profiles. Utilization of glycerolipid substrates and sensitivity to lipase inhibitors suggest that BAT5 is a genuine lipase with preference for long-chain unsaturated MAGs and could in this capacity regulate glycerolipid metabolism in vivo as well. This preliminary SAR data should pave the way towards increasingly potent and BAT5-selective inhibitors.Juha R SavinainenJayendra Z PatelTeija ParkkariDina Navia-PaldaniusJoona J T MarjamaaTuomo LaitinenTapio NevalainenJarmo T LaitinenPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 10, p e109869 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Juha R Savinainen Jayendra Z Patel Teija Parkkari Dina Navia-Paldanius Joona J T Marjamaa Tuomo Laitinen Tapio Nevalainen Jarmo T Laitinen Biochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).
description	<h4>Background</h4>Human lymphocyte antigen B-associated transcript 5 (BAT5, also known as ABHD16A) is a poorly characterized 63 kDa protein belonging to the α/β-hydrolase domain (ABHD) containing family of metabolic serine hydrolases. Its natural substrates and biochemical properties are unknown.<h4>Methodology/principal findings</h4>Amino acid sequence comparison between seven mammalian BAT5 orthologs revealed that the overall primary structure was highly (≥95%) conserved. Activity-based protein profiling (ABPP) confirmed successful generation of catalytically active human (h) and mouse (m) BAT5 in HEK293 cells, enabling further biochemical characterization. A sensitive fluorescent glycerol assay reported hBAT5-mediated hydrolysis of medium-chain saturated (C14:0), long-chain unsaturated (C18:1, C18:2, C20:4) monoacylglycerols (MAGs) and 15-deoxy-Δ12,14-prostaglandin J2-2-glycerol ester (15d-PGJ2-G). In contrast, hBAT5 possessed only marginal diacylglycerol (DAG), triacylglycerol (TAG), or lysophospholipase activity. The best MAG substrates were 1-linoleylglycerol (1-LG) and 15d-PGJ2-G, both exhibiting low-micromolar Km values. BAT5 had a neutral pH optimum and showed preference for the 1(3)- vs. 2-isomers of MAGs C18:1, C18:2 and C20:4. Inhibitor profiling revealed that β-lactone-based lipase inhibitors were nanomolar inhibitors of hBAT5 activity (palmostatin B > tetrahydrolipstatin > ebelactone A). Moreover, the hormone-sensitive lipase inhibitor C7600 (5-methoxy-3-(4-phenoxyphenyl)-3H-[1], [3], [4]oxadiazol-2-one) was identified as a highly potent inhibitor (IC50 8.3 nM). Phenyl and benzyl substituted analogs of C7600 with increased BAT5 selectivity were synthesized and a preliminary SAR analysis was conducted to obtain initial insights into the active site dimensions.<h4>Conclusions/significance</h4>This study provides an initial characterization of BAT5 activity, unveiling the biochemical and pharmacological properties with in vitro substrate preferences and inhibitor profiles. Utilization of glycerolipid substrates and sensitivity to lipase inhibitors suggest that BAT5 is a genuine lipase with preference for long-chain unsaturated MAGs and could in this capacity regulate glycerolipid metabolism in vivo as well. This preliminary SAR data should pave the way towards increasingly potent and BAT5-selective inhibitors.
format	article
author	Juha R Savinainen Jayendra Z Patel Teija Parkkari Dina Navia-Paldanius Joona J T Marjamaa Tuomo Laitinen Tapio Nevalainen Jarmo T Laitinen
author_facet	Juha R Savinainen Jayendra Z Patel Teija Parkkari Dina Navia-Paldanius Joona J T Marjamaa Tuomo Laitinen Tapio Nevalainen Jarmo T Laitinen
author_sort	Juha R Savinainen
title	Biochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).
title_short	Biochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).
title_full	Biochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).
title_fullStr	Biochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).
title_full_unstemmed	Biochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).
title_sort	biochemical and pharmacological characterization of the human lymphocyte antigen b-associated transcript 5 (bat5/abhd16a).
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/7badfbd1be224b0b86ab6594956a58a2
work_keys_str_mv	AT juharsavinainen biochemicalandpharmacologicalcharacterizationofthehumanlymphocyteantigenbassociatedtranscript5bat5abhd16a AT jayendrazpatel biochemicalandpharmacologicalcharacterizationofthehumanlymphocyteantigenbassociatedtranscript5bat5abhd16a AT teijaparkkari biochemicalandpharmacologicalcharacterizationofthehumanlymphocyteantigenbassociatedtranscript5bat5abhd16a AT dinanaviapaldanius biochemicalandpharmacologicalcharacterizationofthehumanlymphocyteantigenbassociatedtranscript5bat5abhd16a AT joonajtmarjamaa biochemicalandpharmacologicalcharacterizationofthehumanlymphocyteantigenbassociatedtranscript5bat5abhd16a AT tuomolaitinen biochemicalandpharmacologicalcharacterizationofthehumanlymphocyteantigenbassociatedtranscript5bat5abhd16a AT tapionevalainen biochemicalandpharmacologicalcharacterizationofthehumanlymphocyteantigenbassociatedtranscript5bat5abhd16a AT jarmotlaitinen biochemicalandpharmacologicalcharacterizationofthehumanlymphocyteantigenbassociatedtranscript5bat5abhd16a
_version_	1718414349794541568

Biochemical and pharmacological characterization of the human lymphocyte antigen B-associated transcript 5 (BAT5/ABHD16A).

Ejemplares similares