Publikationer från Malmö universitet
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The Production of Matchout-Deuterated Cholesterol and the Study of Bilayer-Cholesterol Interactions
Malmö universitet, Fakulteten för hälsa och samhälle (HS), Institutionen för biomedicinsk vetenskap (BMV). Malmö universitet, Biofilms Research Center for Biointerfaces.ORCID-id: 0000-0003-3458-887X
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2019 (Engelska)Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 9, nr 1, artikel-id 5118Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The deuteration of biomolecules provides advanced opportunities for neutron scattering studies. For low resolution studies using techniques such as small-angle neutron scattering and neutron reflection, the level of deuteration of a sample can be varied to match the scattering length density of a specific DO/HO solvent mixture. This can be of major value in structural studies where specific regions of a complex system can be highlighted, and others rendered invisible. This is especially useful in analyses of the structure and dynamics of membrane components. In mammalian membranes, the presence of cholesterol is crucial in modulating the properties of lipids and in their interaction with proteins. Here, a protocol is described for the production of partially deuterated cholesterol which has a neutron scattering length density that matches that of 100% DO solvent (hereby named matchout cholesterol). The level of deuteration was determined by mass spectrometry and nuclear magnetic resonance. The cholesterol match-point was verified experimentally using small angle neutron scattering. The matchout cholesterol was used to investigate the incorporation of cholesterol in various phosphatidylcholine supported lipid bilayers by neutron reflectometry. The study included both saturated and unsaturated lipids, as well as lipids with varying chain lengths. It was found that cholesterol is distributed asymmetrically within the bilayer, positioned closer to the headgroups of the lipids than to the middle of the tail core, regardless of the phosphatidylcholine species.

Ort, förlag, år, upplaga, sidor
Springer Nature, 2019. Vol. 9, nr 1, artikel-id 5118
Nationell ämneskategori
Medicin och hälsovetenskap
Identifikatorer
URN: urn:nbn:se:mau:diva-14810DOI: 10.1038/s41598-019-41439-zISI: 000462298600005PubMedID: 30914734Lokalt ID: 29615OAI: oai:DiVA.org:mau-14810DiVA, id: diva2:1418331
Tillgänglig från: 2020-03-30 Skapad: 2020-03-30 Senast uppdaterad: 2023-10-05Bibliografiskt granskad
Ingår i avhandling
1. Model Membranes and Their Interactions with Native and Artificial Lipoproteins
Öppna denna publikation i ny flik eller fönster >>Model Membranes and Their Interactions with Native and Artificial Lipoproteins
2020 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Atherosclerosis arises from build-up of plaque in the blood, can result in cardiovascular disease and is the largest killer in the west. Low- and high-density lipoproteins are involved in the disease development by depositing and removing lipids to and from artery walls. These processes are complex and not fully understood however, therefore determining the specific roles of the components involved is of fundamental importance in the treatment of the disease.

The work presented in this thesis investigates the production of recombinant tailor-deuterated cholesterol, the structure of cholesterol-containing model membranes and interactions of both native and reconstituted lipoproteins with model membranes. Deuteration is commonly used in neutron scattering for biological samples to provide highly important contrast and the complexity of the native lipoproteins leads to the use of more simple model systems where the compositions can be altered and investigated systematically.

A protocol was developed to produce matchout-deuterated cholesterol for use in neutron scattering studies, as cholesterol is a hugely important component in membranes. The verification of the matchpoint of cholesterol was determined by small-angle neutron scattering and the localisation of cholesterol in model membranes was determined through the use of neutron reflectometry. The interactions of the native and reconstituted lipoproteins with model membranes were also followed by neutron reflectometry, while the structural characterisation of the reconstituted lipoproteins was carried out by small-angle scattering.

Ort, förlag, år, upplaga, sidor
Malmö: Malmö universitet, 2020. s. 74
Serie
Malmö University Health and Society Dissertations, ISSN 1653-5383 ; 2020:4
Nyckelord
Model Cell Membranes, Cholesterol, Lipoproteins, Apolipoprotein E, Lipid Exchange, Reconstituted HDL, Atherosclerosis, Deuteration, Neutron Reflectometry, Small-Angle Neutron Scattering
Nationell ämneskategori
Fysikalisk kemi Biokemi och molekylärbiologi
Identifikatorer
urn:nbn:se:mau:diva-36867 (URN)10.24834/isbn.9789178771325 (DOI)978-91-7877-131-8 (ISBN)978-91-7877-132-5 (ISBN)
Disputation
2020-11-25, Orkanen Hall D138, Nordenskiöldsgatan 10, Malmö, 13:00 (Engelska)
Opponent
Handledare
Tillgänglig från: 2020-11-18 Skapad: 2020-11-18 Senast uppdaterad: 2023-10-05Bibliografiskt granskad

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Waldie, SarahMaric, SelmaCárdenas, Marité

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Institutionen för biomedicinsk vetenskap (BMV)Biofilms Research Center for Biointerfaces
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