Structure and mechanism of human diacylglycerol O-acyltransferase 1

Wang, Lie; Qian, Hui‐Fen; Nian, Yin; Han, Yimo; Ren, Zhenning; Zhang, Hanzhi; Hu, Liya; Prasad, B. V. Venkataram; Laganowsky, Arthur; Yan, Nieng; Zhou, Ming

doi:10.1038/s41586-020-2280-2

Cited by 86 publications

(113 citation statements)

References 69 publications

(57 reference statements)

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“…The absolutely conserved histidine residue that serves as one of the defining characteristics of MBOAT family members (H338 in GOAT) has been suggested to act as a general base in the acylation reactions catalyzed by these enzymes. (19,23,25,28,(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41) While this catalytic role has not been conclusively demonstrated in any MBOAT, the dependence of ligand 15 uptake on the presence on H338 in GOAT supports a direct interaction between the acylation site serine in ghrelin and this conserved histidine (Figure 2b). The enhanced binding affinity of ghrelin ligands with amine modifications at the serine acylation site likely arises from re-apportionment of the transition-state stabilization energy from the serine -histidine hydrogen bond/general base interaction during acyl transfer to ground-state binding enhancement from the amine/ammonium -histidine interaction in ligand 15.…”

Section: Discussionmentioning

confidence: 99%

A remarkably specific ligand reveals ghrelinO-acyltransferase interacts with extracellular peptides and exhibits unexpected cellular localization for a secretory pathway enzyme

Campana

Davis

Cleverdon

et al. 2021

Preprint

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Ghrelin O-acyltransferase (GOAT) plays a central role in the maturation and activation of the peptide hormone ghrelin, which performs a wide range of endocrinological signaling roles. Using a tight-binding fluorescent ghrelin-derived peptide designed for high selectivity for GOAT over the ghrelin receptor GHS-R1a, we demonstrate that GOAT interacts with extracellular ghrelin and facilitates ligand cell internalization in both transfected cells and prostate cancer cells endogenously expressing GOAT. Coupled with enzyme mutagenesis, ligand uptake studies provide the first direct evidence supporting interaction of the putative histidine general base within GOAT with the ghrelin peptide acylation site. Our work provides a new understanding of GOAT's catalytic mechanism, establishes a key step required for autocrine/paracrine ghrelin signaling involving local reacylation by GOAT, and raises the possibility that other peptide hormones may exhibit similar complexity in their intercellular and organismal-level signaling pathways.

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Section: Discussionmentioning

confidence: 99%

A remarkably specific ligand reveals ghrelinO-acyltransferase interacts with extracellular peptides and exhibits unexpected cellular localization for a secretory pathway enzyme

Campana

Davis

Cleverdon

et al. 2021

Preprint

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“…Wei et al (2017) detected that NoDGAT1A expression in C. reinhardtii UVM4 had no effect on TAG accumulation under nitrogen-replete condition but TAG enhancement was observed under nitrogen-depleted condition has been detected using AutoDock [64,65]. However, compare to the presence of cryo-electron microscopy structure of human DGAT1 in complex with an oleoyl-CoA substrate [66,67], the absence 3D structures of DGAT2s hampers the accuracy of AudoDock results.…”

Section: Discussionmentioning

confidence: 99%

Characterization of type-2 diacylglycerol acyltransferases in Haematococcus lacustris reveals their functions and engineering potential in triacylglycerol biosynthesis and possible roles in astaxanthin esterification

Cui¹,

Zhao²,

Xu³

et al. 2020

Preprint

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Background: Haematococcus lacustris is an ideal source of astaxanthin (AST) which is stored at oil bodies containing esterified AST (EAST) and triacylglycerol (TAG). Diacylglycerol acyltransferases (DGATs) catalyze the last step of the acyl-CoA-dependent TAG biosynthesis and are also considered as the crucial enzymes involving in EAST biosynthesis in H. lacustris. Previous studies have identified four putative DGAT2 encoding genes in H. lacustris and only the HpDGAT2D allowed the recovery of TAG biosynthesis, but the engineering potential of HpDGAT2s in TAG biosynthesis, especially possible roles in AST esterification, remains ambiguous.Results: Five putative DGAT2 genes (HpDGAT2A, 2B, 2C, 2D, and 2E) were identified in H. lacustris. Transcription analysis showed that the expression levels of HpDGAT2A, 2B, and 2E genes markedly increased under high light and nitrogen deficient conditions with distinct patterns, which led to significant TAG and EAST accumulation. Functional complementation demonstrated HpDGAT2A, 2B, 2D, and 2E had the capability to restore TAG synthesis in a TAG-deficient yeast strain (H1246) with the large difference in enzymatic activity. Fatty acids (FAs) profiles assays revealed that HpDGAT2A, 2D, and 2E except for 2B preferred monounsaturated fatty acyl-CoA (MUFA) for TAG synthesis in yeast cells, and also showed polyunsaturated fatty acyl-CoA (PUFA) preference by feeding strategy. The over-expression of HpDGAT2D in Arabidopsis thaliana and Chlamydomonas reinhardtii significantly increased the TAG content and obviously promoted the MUFA and PUFA contents. Interestingly, molecular docking analysis implied that HpDGAT2s structures contained AST binding sites, which provides strong evidence for AST esterification function in H. lacustris.Conclusions: Our study represents a pioneering work on the characterization of HpDGAT2s by systematically integrating expression pattern, AST/TAG accumulation, functional complementation, molecular docking, and over-expression in yeast, plants, and algae. These results (1) update the gene models of HpDGAT2s, (2) prove TAG biosynthesis capacity of HpDGAT2s, (3) show the strong preference for MUFA and PUFA, (4) offer target genes to modulate TAG biosynthesis by genetic engineering method, and (5) provide new evidence for HpDGAT2s roles in AST esterification.

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“…Recently, binding sites of potential protease inhibitors of COVID-19 and 3D structure of COVID-19 has been detected using AutoDock [64,65]. However, compare to the presence of cryoelectron microscopy structure of human DGAT1 in complex with an oleoyl-CoA substrate [66,67], the absence 3D structures of DGAT2s hampers the accuracy of AudoDock results.…”

Section: Discussionmentioning

confidence: 99%

Characterization of type-2 diacylglycerol acyltransferases in Haematococcus lacustris reveals their functions and engineering potential in triacylglycerol biosynthesis and possible roles in astaxanthin esterification

Cui

Zhao

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Haematococcus lacustris is an ideal source of astaxanthin (AST) which is stored at oil bodies containing esterified AST (EAST) and triacylglycerol (TAG). Diacylglycerol acyltransferases (DGATs) catalyze the last step of the acyl-CoA-dependent TAG biosynthesis and are also considered as the crucial enzymes involving in EAST biosynthesis in H. lacustris. Previous studies have identified four putative DGAT2 encoding genes in H. lacustris and only the HpDGAT2D allowed the recovery of TAG biosynthesis, but the engineering potential of HpDGAT2s in TAG biosynthesis, especially possible roles in AST esterification, remains ambiguous.Results: Five putative DGAT2 genes (HpDGAT2A, HpDGAT2B, HpDGAT2C, HpDGAT2D, and HpDGAT2E) were identified in H. lacustris. Transcription analysis showed that the expression levels of HpDGAT2A, HpDGAT2D, and HpDGAT2E genes markedly increased under high light and nitrogen deficient conditions with distinct patterns, which led to significant TAG and EAST accumulation. Functional complementation demonstrated HpDGAT2A, HpDGAT2B, HpDGAT2D, and HpDGAT2E had the capability to restore TAG synthesis in a TAG-deficient yeast strain (H1246) with the large difference in enzymatic activity. Fatty acids (FAs) profiles assays revealed that HpDGAT2A, HpDGAT2D, and HpDGAT2E except for HpDGAT2B preferred monounsaturated fatty acyl-CoAs (MUFAs) for TAG synthesis in yeast cells, and also showed polyunsaturated fatty acyl-CoAs (PUFAs) preference by feeding strategy. The over-expression of HpDGAT2D in Arabidopsis thaliana and Chlamydomonas reinhardtii significantly increased the TAG content and obviously promoted the MUFAs and PUFAs contents. Interestingly, molecular docking analysis implied that HpDGAT2s structures contained AST binding sites, which provides strong evidence for AST esterification function in H. lacustris.Conclusions: Our study represents a pioneering work on the characterization of HpDGAT2s by systematically integrating expression pattern, AST/TAG accumulation, functional complementation, molecular docking, and over-expression in yeast, plants, and algae. These results (1) update the gene models of HpDGAT2s, (2) prove TAG biosynthesis capacity of HpDGAT2s, (3) show the strong preference for MUFAs and PUFAs, (4) offer target genes to modulate TAG biosynthesis by genetic engineering method, and (5) provide new evidence for HpDGAT2s roles in AST esterification.

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Structure and mechanism of human diacylglycerol O-acyltransferase 1

Cited by 86 publications

References 69 publications

A remarkably specific ligand reveals ghrelinO-acyltransferase interacts with extracellular peptides and exhibits unexpected cellular localization for a secretory pathway enzyme

A remarkably specific ligand reveals ghrelinO-acyltransferase interacts with extracellular peptides and exhibits unexpected cellular localization for a secretory pathway enzyme

Characterization of type-2 diacylglycerol acyltransferases in Haematococcus lacustris reveals their functions and engineering potential in triacylglycerol biosynthesis and possible roles in astaxanthin esterification

Characterization of type-2 diacylglycerol acyltransferases in Haematococcus lacustris reveals their functions and engineering potential in triacylglycerol biosynthesis and possible roles in astaxanthin esterification

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