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2.A.50 The Glycerol Uptake (GUP) or Membrane-bound Acyl Transferase (MBOAT) Family

Yeast can use glycerol both as a carbon source and as an osmolyte. In Saccharomyces cerevisiae, glycerol has been reported to be actively taken up via two electrogenic H+ symporters, GUP1 (YGL084c) and GUP2 (YPL189w). These proteins were the first characterized members in a family of polytopic integral membrane proteins from fungi, animals and bacteria. Several of the bacterial proteins have been implicated in extracellular polysaccharide modification. One such protein, DltB (TC# 2.A.50.2.1; P39580) has been implicated in the transport of activated D-alanine across the bacterial cytoplasmic membrane for biosynthesis of D-alanine lipoteichoic acid. It is possible that proteins implicated in polysaccharide modification serve a similar role. Most of these proteins are of between 450 and 610 aas in length and exhibit 8-10 putative TMSs. One protein (spO09758) from Schizosaccharomyces pombe is half sized, having only 231aas with 4-5 putative TMSs. These proteins are members of the membrane-bound O acyl transferase (MBOAT) family. 

The Bacillus subtilis dlt operon (D-alanyl-lipoteichoic acid) is responsible for D-alanine esterification of both lipoteichoic acid (LTA) and wall teichoic acid (WTA). The dlt operon contains five genes, dltA-dltE. Insertional inactivation of dltA-dltD results in complete absence of D-alanine in both LTA and WTA. Based on protein sequence similarity with the Lactobacillus casei dlt gene products (Heaton and Neuhaus, 1992), dltA may encode the D-alanine-D-alanyl carrier protein ligase (Dcl) and dltC the D-alanyl carrier protein (Dcp). The products of dltB and dltD are concerned with the transport of activated D-alanine through the membrane and the final incorporation of D-alanine into LTA. The hydropathy profiles of the DltB and DltD suggest a transmembrane location for the former and an amino-terminal signal peptide for the latter (Perego et al., 1995).

Gup1 homologues of S. cerevisiae and Trypanosoma brucei are glycosyl phosphatidylinositol (GPI) remodelases (Jaquenoud et al., 2008; Ghugtyal et al., 2007). Lipids of Trypanosoma brucei undergo lipid remodelling, whereby longer fatty acids on the glycerol are replaced by myristate (C14:0). A similar process occurs on GPI proteins of Saccharomyces cerevisiae where Per1p first deacylates and Gup1p subsequently reacylates the anchor lipid, thus replacing a shorter fatty acid by C26:0 (Ghugtyal et al., 2007). Heterologous expression of the GUP1 homologue of T. brucei in Δgup1 yeast cells partially normalizes the Δgup1 phenotype and restores the transfer of labelled fatty acids from Coenzyme A to lyso-GPI proteins. Gup1p from T. brucei (tbGup1p) strongly prefers C14:0 and C12:0 over C16:0 and C18:0, whereas yeast Gup1p strongly prefers C16:0 and C18:0. This acyl specificity of tbGup1p closely matches the reported specificity of the reacylation of free lyso-GPI lipids in microsomes of T. brucei. Depletion of tbGup1p in trypanosomes by RNAi drastically reduces the rate of myristate incorporation into the sn-2 position of lyso-GPI lipids. Thus, tbGup1p is involved in the addition of myristate to sn-2 during GPI remodelling in T. brucei and can account for the fatty acid specificity of this process. tbGup1p can act on GPI proteins as well as on GPI lipids. These results and others put into doubt the original claim that GUP1 and GUP2 are glycerol transporters (Bleve et al., 2005; Bosson et al., 2006; Ferreira et al., 2006).

Mammalian glycerol uptake/transporter 1 (Gup1), a homolog of Saccharomyces cerevisiae Gup1, is predicted to be a member of the membrane-bound O-acyltransferase family and is highly homologous to mammalian hedgehog acyltransferase, known as Skn, the homolog of the Drosophila skinny hedgehog gene product. Although mammalian Gup1 has a sequence conserved among the membrane-bound O-acyltransferase family, the histidine residue in the motif that is indispensable to the acyltransferase activity of the family has been replaced with leucine. Abe et al. (2007) cloned Gup1 cDNA from adult mouse lung and examined whether it is involved in the regulation of N-terminal palmitoylation of Sonic hedgehog (Shh). Subcellular localization of mouse Gup1 was indistinguishable from that of mouse Skn. Gup1 and Skn co-localized with endoplasmic reticulum markers, suggesting that these two molecules interact with overlapped targets. Ectopic expression of Gup1 with full-length Shh in cells lacking endogenous Skn showed no hedgehog acyltransferase activity. On the other hand, Gup1 interfered with the palmitoylation of Shh catalyzed by endogenous Skn in COS7 and NSC34. Gup1 may thus be a negative regulator of N-terminal palmitoylation of Shh and may contribute to the variety of biological actions of Shh (Abe et al., 2007). MBOAT7 is anchared to endomembranes by a 6 TMS domain, and it functions in remodeling the acyl chain compositions of endomembranes (Caddeo et al. 2019).

Membrane-bound O-acyltransferases (MBOATs) are a superfamily of integral transmembrane enzymes that are found in all kingdoms of life. In bacteria, MBOATs modify protective cell-surface polymers while in vertebrates, some MBOAT enzymes - such as acyl-coenzyme A:cholesterol acyltransferase and diacylglycerol acyltransferase 1 - are responsible for lipid biosynthesis or phospholipid remodelling. Other MBOATs, including porcupine, hedgehog acyltransferase and ghrelin acyltransferase, catalyse essential lipid modifications of secreted proteins such as Wnt, hedgehog and ghrelin, respectively. Many MBOAT proteins are important drug targets. Ma et al. 2018 presented crystal structures of DltB, an MBOAT responsible for the D-alanylation of cell-wall teichoic acid, both alone and in complex with the D-alanyl donor protein DltC. DltB contains a ring of 11 peripheral transmembrane helices, which shield a highly conserved extracellular structural funnel extending into the middle of the lipid bilayer. The conserved catalytic histidine residue is located at the bottom of this funnel and is connected to the intracellular DltC through a narrow tunnel. Mutation of either the catalytic histidine or the DltC-binding site of DltB abolishes the D-alanylation of lipoteichoic acid and sensitizes B. subtilis to cell-wall stress, which suggests cross-membrane catalysis involving the tunnel. Structure-guided sequence comparisons among DltB and vertebrate MBOATs reveal a conserved structural core and suggests that MBOATs from different organisms have similar catalytic mechanisms. 

The protein, porcupine (2.A.50.3.1 and 2), is a  palmiteoyl transferase, known to acylate regulatory proteins such as Wnt.  It also regulates certain channel proteins.  For example it regulates the AMPA/TARP complex in a TARP-dependent manner (Kato and Witkin 2018).

Diacylglycerol O-acyltransferase 1, DGAT1 or AGRP1, of 488 aas and 9 TMSs, has been structurally solved by cryoEM (Wang et al. 2020). It synthesizes triacylglycerides and is required for dietary fat absorption and fat storage in humans. DGAT1 belongs to the membrane-bound O-acyltransferase (MBOAT) superfamily, members of which are found in all kingdoms of life and are involved in the acylation of lipids and proteins. Wang et al. 2020 addressed how human DGAT1 and other mammalian members of the MBOAT family recognize their substrates and catalyse their reactions. They revealed the structure of human DGAT1 in complex with oleoyl-CoA. Each DGAT1 protomer has nine TMSs, eight of which form a conserved structural fold, the MBOAT fold. It forms a hollow chamber in the membrane that encloses highly conserved catalytic residues. The chamber has separate entrances for each of the two substrates, fatty acyl-CoA and diacylglycerol. DGAT1 exists as either a homodimer or a homotetramer, and the two forms have similar enzymatic activities. The N terminus of DGAT1 interacts with the neighbouring protomer, and these interactions are required for enzymatic activity (Wang et al. 2020).


The transport reaction proposed to be catalyzed by GUP1 is:

Glycerol (out) + H+ (out) → glycerol (in) + H+ (in)

The transport reaction proposed to be catalyzed by DltB is:

DctC-alanine (in) + Teichoic acid (out) → Alanyl-teichoic acid (out) + DctC (in)

References associated with 2.A.50 family:

Abe, Y., Y. Kita, and T. Niikura. (2008). Mammalian Gup1, a homolog of Saccharomyces cerevisiae glycerol uptake/transporter 1, acts as a negative regulator for N-terminal palmitoylation of Sonic hedgehog. F.E.B.S.J. 275: 318-331. 18081866
Bleve, G., G. Zacheo, M.S. Cappello, F. Dellaglio, and F. Grieco. (2005). Subcellular localization and functional expression of the glycerol uptake protein 1 (GUP1) of Saccharomyces cerevisiae tagged with green fluorescent protein. Biochem. J. 390: 145-155. 15813700
Bosson, R., M. Jaquenoud, and A. Conzelmann. (2006). GUP1 of Saccharomyces cerevisiae encodes an O-acyltransferase involved in remodeling of the GPI anchor. Mol. Biol. Cell 17: 2636-2645. 16597698
Caddeo, A., O. Jamialahmadi, G. Solinas, A. Pujia, R.M. Mancina, P. Pingitore, and S. Romeo. (2019). MBOAT7 is anchored to endomembranes by six transmembrane domains. J Struct Biol. [Epub: Ahead of Print] 30959108
Campaña, M.B., F.J. Irudayanathan, T.R. Davis, K.R. McGovern-Gooch, R. Loftus, M. Ashkar, N. Escoffery, M. Navarro, M.A. Sieburg, S. Nangia, and J.L. Hougland. (2019). The ghrelin -acyltransferase structure reveals a catalytic channel for transmembrane hormone acylation. J. Biol. Chem. [Epub: Ahead of Print] 31413115
Chamoun, Z., R.K. Mann, D. Nellen, D.P. von Kessler, M. Bellotto, P.A. Beachy, and K. Basler. (2001). Skinny hedgehog, an acyltransferase required for palmitoylation and activity of the hedgehog signal. Science 293: 2080-2084. 11486055
Coupland, C.E., S.A. Andrei, T.B. Ansell, L. Carrique, P. Kumar, L. Sefer, R.A. Schwab, E.F.X. Byrne, E. Pardon, J. Steyaert, A.I. Magee, T. Lanyon-Hogg, M.S.P. Sansom, E.W. Tate, and C. Siebold. (2021). Structure, mechanism, and inhibition of Hedgehog acyltransferase. Mol. Cell 81: 5025-5038.e10. 34890564
Di Sessa, A., S. Guarino, A.P. Passaro, L. Liguori, G.R. Umano, G. Cirillo, E. Miraglia Del Giudice, and P. Marzuillo. (2021). NAFLD and renal function in children: is there a genetic link? Expert Rev Gastroenterol Hepatol 1-10. [Epub: Ahead of Print] 33851883
Ferreira, C., S. Silva, F. van Voorst, C. Aguiar, M.C. Kielland-Brandt, A. Brandt, and C. Lucas. (2006). Absence of Gup1p in Saccharomyces cerevisiae results in defective cell wall composition, assembly, stability and morphology. FEMS Yeast Res. 6: 1027-1038. 17042752
Ghugtyal, V., C. Vionnet, C. Roubaty, and A. Conzelmann. (2007). CWH43 is required for the introduction of ceramides into GPI anchors in Saccharomyces cerevisiae. Mol. Microbiol. 65: 1493-1502. 17714445
Gutierrez, J.A., P.J. Solenberg, D.R. Perkins, J.A. Willency, M.D. Knierman, Z. Jin, D.R. Witcher, S. Luo, J.E. Onyia, and J.E. Hale. (2008). Ghrelin octanoylation mediated by an orphan lipid transferase. Proc. Natl. Acad. Sci. USA 105: 6320-6325. 18443287
Heaton, M.P. and F.C. Neuhaus. (1992). Biosynthesis of D-alanyl-lipoteichoic acid: cloning, nucleotide sequence, and expression of the Lactobacillus casei gene for the D-alanine-activating enzyme. J. Bacteriol. 174: 4707-4717. 1385594
Helsley, R.N., V. Venkateshwari, A.L. Brown, A.D. Gromovsky, R.C. Schugar, I. Ramachandiran, K. Fung, M.N. Kabbany, R. Banerjee, C.K. Neumann, C. Finney, P. Pathak, O. Danny, L.J. Osborn, W. Massey, R. Zhang, A. Kadam, B.E. Sansbury, C. Pan, J. Sacks, R.G. Lee, R.M. Crooke, M.J. Graham, M.E. Lemieux, V. Gogonea, J.P. Kirwan, D.S. Allende, M. Civelek, P.L. Fox, L.L. Rudel, A.J. Lusis, M. Spite, and J.M. Brown. (2019). Obesity-linked suppression of membrane-bound -Acyltransferase 7 (MBOAT7) drives non-alcoholic fatty liver disease. Elife 8:. [Epub: Ahead of Print] 31621579
Holst, B., C. Lunde, F. Lages, R. Oliveira, C. Lucas and M.C. Kielland-Brandt (2000). GUP1 and its close homologue GUP2, encoding multimembrane-spaning proteins involved in active glycerol uptake in Saccharomyces cerevisiae. Mol. Microbiol. 37: 108-124. 10931309
Jaquenoud, M., M. Pagac, A. Signorell, M. Benghezal, J. Jelk, P. Bütikofer, and A. Conzelmann. (2008). The Gup1 homologue of Trypanosoma brucei is a GPI glycosylphosphatidylinositol remodelase. Mol. Microbiol. 67: 202-212. 18036137
Jeppson, S., H. Mattisson, K. Demski, and I. Lager. (2020). A predicted transmembrane region in plant diacylglycerol acyltransferase 2 regulates specificity towards very long chain acyl-CoAs. J. Biol. Chem. [Epub: Ahead of Print] 32873712
Jiang, Y., T.L. Benz, and S.B. Long. (2021). Substrate and product complexes reveal mechanisms of Hedgehog acylation by HHAT. Science 372: 1215-1219. 34112694
Lee, H.C., T. Inoue, R. Imae, N. Kono, S. Shirae, S. Matsuda, K. Gengyo-Ando, S. Mitani, and H. Arai. (2008). Caenorhabditis elegans mboa-7, a member of the MBOAT family, is required for selective incorporation of polyunsaturated fatty acids into phosphatidylinositol. Mol. Biol. Cell 19: 1174-1184. 18094042
Lee, J.D. and J.E. Treisman. (2001). Sightless has homology to transmembrane acyltransferases and is required to generate active Hedgehog protein. Curr. Biol. 11: 1147-1152. 11509241
Longo, M., M. Meroni, E. Paolini, V. Erconi, F. Carli, F. Fortunato, D. Ronchi, R. Piciotti, S. Sabatini, C. Macchi, A. Alisi, L. Miele, G. Soardo, G.P. Comi, L. Valenti, M. Ruscica, A.L. Fracanzani, A. Gastaldelli, and P. Dongiovanni. (2021). TM6SF2/PNPLA3/MBOAT7 Loss-of-Function Genetic Variants Impact on NAFLD Development and Progression Both in Patients and in In Vitro Models. Cell Mol Gastroenterol Hepatol 13: 759-788. [Epub: Ahead of Print] 34823063
Lucas, C., C. Ferreira, G. Cazzanelli, R. Franco-Duarte, and J. Tulha. (2016). Yeast Gup1(2) Proteins Are Homologues of the Hedgehog Morphogens Acyltransferases HHAT(L): Facts and Implications. J Dev Biol 4:. 29615596
Ma, D., Z. Wang, C.N. Merrikh, K.S. Lang, P. Lu, X. Li, H. Merrikh, Z. Rao, and W. Xu. (2018). Crystal structure of a membrane-bound O-acyltransferase. Nature 562: 286-290. 30283133
Ma, Z., J.M. Onorato, L. Chen, D.W. Nelson, C.E. Yen, and D. Cheng. (2017). Synthesis of neutral ether lipid monoalkyl-diacylglycerol by lipid acyltransferases. J Lipid Res 58: 1091-1099. 28420705
Matsuda, S., T. Inoue, H.C. Lee, N. Kono, F. Tanaka, K. Gengyo-Ando, S. Mitani, and H. Arai. (2008). Member of the membrane-bound O-acyltransferase (MBOAT) family encodes a lysophospholipid acyltransferase with broad substrate specificity. Genes Cells 13: 879-888. 18782225
Perego, M., P. Glaser, A. Minutello, M.A. Strauch, K. Leopold, and W. Fischer W. (1995). Incorporation of D-alanine into lipoteichoic acid and wall teichoic acid in Bacillus subtilis. Identification of genes and regulation. J. Biol. Chem. 270: 15598-15606. 7797557
Wang, L., H. Qian, Y. Nian, Y. Han, Z. Ren, H. Zhang, L. Hu, B.V.V. Prasad, A. Laganowsky, N. Yan, and M. Zhou. (2020). Structure and mechanism of human diacylglycerol O-acyltransferase 1. Nature 581: 329-332. 32433610