Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate

Roberto Stock, Jonathan R. Brewer, Kerstin Wagner, Blanca Ramos-Cerrillo, Lars Duelund, Kit Drescher Jernshøj, Lars Folke Olsen, Luis Bagatolli

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1) ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate) can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2) the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3) in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes.
OriginalsprogEngelsk
TidsskriftPLOS ONE
Vol/bind7
Udgave nummer4
Sider (fra-til)1-15 (e-36003)
Antal sider15
ISSN1932-6203
StatusUdgivet - 25. apr. 2012

Fingeraftryk

ceramides
sphingomyelins
Sphingomyelins
phosphates
Membranes
Loxosceles
Enzyme kinetics
Unilamellar Liposomes
Enzyme activity
Enzymes
Araneae
fluorescence emission spectroscopy
Spider Venoms
enzyme activity
Membrane structures
enzyme kinetics
Fluorescence microscopy
Fluorescence spectroscopy
venoms
light scattering

Citer dette

Stock, R., Brewer, J. R., Wagner, K., Ramos-Cerrillo, B., Duelund, L., Jernshøj, K. D., ... Bagatolli, L. (2012). Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate. PLOS ONE, 7(4), 1-15 (e-36003).
Stock, Roberto ; Brewer, Jonathan R. ; Wagner, Kerstin ; Ramos-Cerrillo, Blanca ; Duelund, Lars ; Jernshøj, Kit Drescher ; Olsen, Lars Folke ; Bagatolli, Luis. / Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate. I: PLOS ONE. 2012 ; Bind 7, Nr. 4. s. 1-15 (e-36003).
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title = "Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate",
abstract = "The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1) ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate) can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2) the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3) in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes.",
author = "Roberto Stock and Brewer, {Jonathan R.} and Kerstin Wagner and Blanca Ramos-Cerrillo and Lars Duelund and Jernsh{\o}j, {Kit Drescher} and Olsen, {Lars Folke} and Luis Bagatolli",
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Stock, R, Brewer, JR, Wagner, K, Ramos-Cerrillo, B, Duelund, L, Jernshøj, KD, Olsen, LF & Bagatolli, L 2012, 'Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate', PLOS ONE, bind 7, nr. 4, s. 1-15 (e-36003).

Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate. / Stock, Roberto ; Brewer, Jonathan R.; Wagner, Kerstin; Ramos-Cerrillo, Blanca; Duelund, Lars; Jernshøj, Kit Drescher; Olsen, Lars Folke; Bagatolli, Luis.

I: PLOS ONE, Bind 7, Nr. 4, 25.04.2012, s. 1-15 (e-36003).

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate

AU - Stock, Roberto

AU - Brewer, Jonathan R.

AU - Wagner, Kerstin

AU - Ramos-Cerrillo, Blanca

AU - Duelund, Lars

AU - Jernshøj, Kit Drescher

AU - Olsen, Lars Folke

AU - Bagatolli, Luis

PY - 2012/4/25

Y1 - 2012/4/25

N2 - The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1) ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate) can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2) the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3) in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes.

AB - The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1) ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate) can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2) the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3) in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes.

M3 - Journal article

VL - 7

SP - 1-15 (e-36003)

JO - P L o S One

JF - P L o S One

SN - 1932-6203

IS - 4

ER -