Sphingolipid transfer proteins defined by the GLTP-fold

Malinina, Lucy; Simanshu, Dhirendra K.; Zhai, Xiuhong; Samygina, V.R.; Kamlekar, Ravi Kanth; Kenoth, Roopa; Ochoa-Lizarralde, B.; Malakhova, Margarita; Molotkovsky, Julian G.; Patel, Dinshaw J.; Brown, Rhoderick E.

doi:10.1017/s003358351400016x

Cited by 32 publications

(50 citation statements)

References 155 publications

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“…The experiment was started by the addition of 0.083 pmol (1.5 g) g-rGM2AP or 0.073 pmol (1.5 g) g-rGM2AP-His 6 . Increase in NBD-fl uorescence in the acceptor vesicle was measured continuously, as described previously ( 34 ), with a spectrofl uorophotometer (RF-5000; Shimadzu, Düsseldorf, Germany).…”

Section: Lipid Transfer Assay By Förster Resonance Energy Transfermentioning

confidence: 99%

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Membrane lipids regulate ganglioside GM2 catabolism and GM2 activator protein activity

Anheuser

Breiden

Schwarzmann

et al. 2015

Journal of Lipid Research

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For the effective degradation of lysosomal sphingolipids with short oligosaccharide head groups, special lipid binding and transfer proteins (the sphingolipid activator proteins) are needed. The sphingolipid activator proteins consist of the GM2 activator protein (GM2AP) and the saposins (Saps) A-D. They all facilitate the interaction of the membrane-bound substrate with the water-soluble enzyme at the water-membrane interface and differ from sphingolipid transfer proteins of the cytosol ( 6 ). The anionic lysosomal phospholipid, BMP, is also known to dramatically enhance (glyco)sphingolipid hydrolysis by lysosomal hydrolases and sphingolipid activator proteins ( 7-9 ).Even at low pH values (pH <5.0), BMP conveys negative surface charge to luminal lysosomal vesicles ( 10,11 ). This favors an electrostatic binding of the positively charged activator proteins and the polycationic hydrolases, and enhances degradation of (glyco)sphingolipids substantially ( 7,9,10,(12)(13)(14)(15)(16).The GM2AP, which has a hydrophobic binding cavity for lipids and a binding site for ␤ -hexosaminidase A (HexA) ( 17-19 ), arranges the interaction of ganglioside GM2 with HexA and facilitates the formation of a Michaelis-Menten complex with access of the catalytic site to the carbohydrate head group ( 20,21 ). The whole working mechanism, Abstract Ganglioside GM2 is the major lysosomal storage compound of Tay-Sachs disease. It also accumulates in Niemann-Pick disease types A and B with primary storage of SM and with cholesterol in type C. Reconstitution of GM2 catabolism with ␤ -hexosaminidase A and GM2 activator protein (GM2AP) at uncharged liposomal surfaces carrying GM2 as substrate generated only a physiologically irrelevant catabolic rate, even at pH 4.2. However, incorporation of anionic phospholipids into the GM2 carrying liposomes stimulated GM2 hydrolysis more than 10-fold, while the incorporation of plasma membrane stabilizing lipids (SM and cholesterol) generated a strong inhibition of GM2 hydrolysis, even in the presence of anionic phospholipids. Mobilization of membrane lipids by GM2AP was also inhibited in the presence of cholesterol or SM, as revealed by surface plasmon resonance studies. These lipids also reduced the interliposomal transfer rate of 2-NBD-GM1 by GM2AP, as observed in assays using Förster resonance energy transfer. Our data raise major concerns about the usage of recombinant His-tagged GM2AP compared with untagged protein. The former binds more strongly to anionic GM2-carrying liposomal surfaces, increases GM2 hydrolysis, and accelerates intermembrane transfer of 2-NBD-GM1, but does not mobilize membrane lipids. Components of eukaryotic plasma membranes reach the intraendolysosomal vesicles by endocytotic membrane fl ow ( 1 ). Those vesicles have been identifi ed as platforms for lipid and membrane degradation ( 2 ). They differ from the limiting endosomal membrane by the absence of a glycocalix, a lower cholesterol content, and the cumulative occurrence of the anionic bis(monoacylglycero)phosphate (BMP) ...

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Section: Lipid Transfer Assay By Förster Resonance Energy Transfermentioning

confidence: 99%

“…Therefore, g-rGM2AP-His 6 was not used in further FRET measurements. of total fused liposomes were measured.…”

Section: Expression and Purifi Cation Of Recombinant Glycosylated Tamentioning

confidence: 99%

Membrane lipids regulate ganglioside GM2 catabolism and GM2 activator protein activity

Anheuser

Breiden

Schwarzmann

et al. 2015

Journal of Lipid Research

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“…In this case, FRET will occur between the probes distributed in the membrane: any change in the distribution of probes affects the energy transfer efficiency, and thereby changes the fluorescence intensity of the donor. FRET between membrane-bound probes was extensively used to study the lateral organization of membranes [6][7][8] and lipid transporting enzymes (for example, probes used in [9] and [10]). …”

Section: Introductionmentioning

confidence: 99%

3-{4-[(E)-{4-[(E)-Phenyldiazenyl]phenyl}diazenyl]phenoxy}propane-1,2-diol

Chupin

Болдырев

2017

Molbank

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Abstract:Title compound was designed to be a black quencher of pyrene fluorescence. It was made amphiphilic to serve as a membrane-bound probe. The synthesis is a two-step procedure. The first step is a Mitsunobu reaction of [{(phenyldiazenyl)phenyl}diazenyl]phenol with 1,2-O-isopropylideneglycerol. The second step is the cleavage of the isopropylidene protecting group. The title compound has the extinction coefficient 59,000 at λmax = 380 nm. The Forster distance between the title compound and the pyrene was found to be 37.8 Å.

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“…Structural homology modeling of the FAPP2-GLTPH domain suggests membership in the GLTP superfamily (1,2,4), a group of eukaryotic LTPs that selectively transfer SLs between membranes (5)(6)(7)(8)(9)(10)(11) and share a common protein fold first established from human GLTP crystal structure (12). The GLTP fold is composed of eight ␣-helices, organized as a curved two-layer "sandwich" that envelopes the SL aliphatic chains within an internal hydrophobic pocket that is accessed via a cleft-like gate, whereas the SL-specific headgroup-binding site includes a surface localized recognition center (12).…”

mentioning

confidence: 99%

“…By contrast, human GLTP efficiently transfers both complex and simple GSLs (6,7,9), as reviewed in Ref. 11. Crystal structures of different holo forms of human GLTP provide insights into the structural features responsible for the recognition of GalCer, GlcCer, LacCer, and SF.…”

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confidence: 99%

Structural analyses of 4-phosphate adaptor protein 2 yield mechanistic insights into sphingolipid recognition by the glycolipid transfer protein family

Ochoa-Lizarralde

Gao

Popov

et al. 2018

Journal of Biological Chemistry

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The glycolipid transfer protein (GLTP) fold defines a superfamily of eukaryotic proteins that selectively transport sphingolipids (SLs) between membranes. However, the mechanisms determining the protein selectivity for specific glycosphingolipids (GSLs) are unclear. Here, we report the crystal structure of the GLTP homology (GLTPH) domain of human 4-phosphate adaptor protein 2 (FAPP2) bound with -oleoyl-galactosylceramide. Using this domain, FAPP2 transports glucosylceramide from its-Golgi synthesis site to the -Golgi for conversion into complex GSLs. The FAPP2-GLTPH structure revealed an element, termed the ID loop, that controls specificity in the GLTP family. We found that, in accordance with FAPP2 preference for simple GSLs, the ID loop protrudes from behind the SL headgroup-recognition center to mitigate binding by complex GSLs. Mutational analyses including GLTP and FAPP2 chimeras with swapped ID loops supported the proposed restrictive role of the FAPP2 ID loop in GSL selectivity. Comparative analysis revealed distinctly designed ID loops in each GLTP family member. This analysis also disclosed a conserved H-bond triplet that "clasps" both ID-loop ends together to promote structural autonomy and rigidity. The findings indicated that various ID loops work in concert with conserved recognition centers to create different specificities among family members. We also observed four bulky, conserved hydrophobic residues involved in "sensor-like" interactions with lipid chains in protein hydrophobic pockets and FF motifs in GLTP and FAPP2, well-positioned to provide acyl chain-dependent SL selectivity for the hydrophobic pockets. In summary, our study provides mechanistic insights into sphingolipid recognition by the GLTP fold and uncovers the elements involved in this recognition.

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Sphingolipid transfer proteins defined by the GLTP-fold

Cited by 32 publications

References 155 publications

Membrane lipids regulate ganglioside GM2 catabolism and GM2 activator protein activity

Membrane lipids regulate ganglioside GM2 catabolism and GM2 activator protein activity

3-{4-[(E)-{4-[(E)-Phenyldiazenyl]phenyl}diazenyl]phenoxy}propane-1,2-diol

Structural analyses of 4-phosphate adaptor protein 2 yield mechanistic insights into sphingolipid recognition by the glycolipid transfer protein family

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