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Semiconstant-Time P,H-COSY NMR: Analysis of Complex Mixtures of Phospholipids Originating from Helicobacter pylori
Umeå universitet, Medicinska fakulteten, Institutionen för medicinsk kemi och biofysik. (Prof Schleucher)
Umeå universitet, Medicinska fakulteten, Institutionen för medicinsk kemi och biofysik. (Arnqvist)
Umeå universitet, Medicinska fakulteten, Institutionen för medicinsk kemi och biofysik.
Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
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2009 (Engelska)Ingår i: Journal of the American Chemical Society, ISSN 0002-7863, E-ISSN 1520-5126, Vol. 131, nr 40, s. 14150-1Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Lipids play a central role in numerous biological events, ranging from normal physiological processes to host−pathogen interactions. The proposed semiconstant-time 31P,1H−COSY NMR experiment provides identification of known and structural characterization of unknown phospholipids in complex membrane extracts with high sensitivity, based on the combination of their 1H and 31P chemical shifts and coupling patterns. Furthermore, the spectra allow quantification of phospholipid composition. Analysis of the phospholipid composition of Helicobacter pylori, the causative agent of peptic ulcer disease, showed the presence of uncommon phospholipids. This novel NMR approach allows the study of changes in membrane composition in response to biological stimuli and opens up the possibility of identifying soluble phosphorus species in a number of research fields.

Ort, förlag, år, upplaga, sidor
ACS Publications ASAP , 2009. Vol. 131, nr 40, s. 14150-1
Identifikatorer
URN: urn:nbn:se:umu:diva-26035DOI: 10.1021/ja905282hOAI: oai:DiVA.org:umu-26035DiVA, id: diva2:236149
Anmärkning
Publication Date (Web): September 17, 2009Tillgänglig från: 2009-09-21 Skapad: 2009-09-21 Senast uppdaterad: 2018-06-08Bibliografiskt granskad
Ingår i avhandling
1. NMR studies of host-pathogen interactions
Öppna denna publikation i ny flik eller fönster >>NMR studies of host-pathogen interactions
2009 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

This thesis describes the use of Nuclear Magnetic Resonance (NMR) for characterizing two host-pathogen interactions: The behavior of a regulatory RNA of the Hepatitis B virus (HBV) and the attachment of Helicobacter pylori (H. pylori) to the gastric mucosa. NMR is a powerful tool in biomedicine, because molecules ranging from small ligands to biomacromolecules can be studied with atomic resolution. Different NMR experiments are designed to determine structures, or to monitor interactions, folding, stability or motion.

Paper I describes the analysis of the motions of a regulatory RNA of HBV. The NMR structure of the RNA had revealed before that several well-conserved nucleotides adopt multiple conformations. Therefore an analysis of possible underlying motions was undertaken using two different NMR techniques, one of which (off-resonance ROESY) was applied to nucleic acids for the first time. The observed motions suggest an explanation why the structurally poorly defined nucleotides are highly conserved.

In paper II we improved the ROESY NMR experiment, which is used to measure internuclear distances for structure determination of medium-sized molecules. Using a small protein and an organometallic complex as examples, we demonstrated that the new EASY ROESY experiment yields clean spectra that can directly be integrated to derive interatomic distances.

H. pylori, the bacterium involved in peptic ulcer disease and gastric cancer, survives in the harsh acidic environment of the stomach. It possesses many membrane proteins which mediate adherence, raising the question, if their activity is related to membrane composition. In paper III & IV we analyzed therefore the phospholipid composition of H. pylori membranes.

In paper III, an advanced method for the analysis of the phospholipid composition of biological membranes was developed. The two-dimensional semi-constant-time 31P,1H-COSY experiment combines information from phosphorus and hydrogen atoms of phospholipids for their unambiguous identification. Furthermore, the high resolution of the two-dimensional experiment allows the quantification of phospholipids where conventional methods fail.

In paper IV we applied the new experiment to analyze the lipid composition of whole H. pylori cells, their inner and outer membranes, and of vesicles shed by the bacterium. The goal of this study was to characterize the vesicles which are suggested to play a role in the inflammation process. We established that the outer membrane and the vesicles have similar phospholipid compositions, suggesting that the vesicles are largely derived from the outer membrane.

The NMR results presented here elucidate details of molecular systems engaged in pathogenicity, as basis for therapeutic strategies against these pathogens.

Ort, förlag, år, upplaga, sidor
Umeå: , 2009. s. 48
Serie
Umeå University medical dissertations, ISSN 0346-6612 ; 1275
Nyckelord
NMR, regulatory RNA, HBV, H.pylori, phospholipids, ROESY
Nationell ämneskategori
Biofysik
Forskningsämne
biokemi
Identifikatorer
urn:nbn:se:umu:diva-25710 (URN)978-91-7264-816-6 (ISBN)
Distributör:
Medicinsk kemi och biofysik, 901 85, Umeå
Disputation
2009-09-26, KB3A9, plan 3, KBC-huset, Umeå Universität, Umeå, 10:00 (Engelska)
Opponent
Handledare
Tillgänglig från: 2009-09-08 Skapad: 2009-08-31 Senast uppdaterad: 2018-06-08Bibliografiskt granskad
2. Helicobacter pylori outer membrane vesicles and the host-pathogen interaction
Öppna denna publikation i ny flik eller fönster >>Helicobacter pylori outer membrane vesicles and the host-pathogen interaction
2013 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Alternativ titel[sv]
Helicobacter pylori membranvesiklar och interaktioner med värdcellen
Ort, förlag, år, upplaga, sidor
Umeå: Umeå universitet, 2013. s. 56
Serie
Umeå University medical dissertations, ISSN 0346-6612 ; 1559
Nyckelord
H. pylori, outer membrane vesicles, 2D 31P, 1H COSY NMR, phospholipids, OMV proteome, adherence, endocytosis, CagA, OMV-host cell responses, yttermembran vesiklar
Nationell ämneskategori
Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)
Identifikatorer
urn:nbn:se:umu:diva-68744 (URN)978-91-7459-578-9 (ISBN)
Disputation
2013-05-17, KB3B1, KBC-huset, Umeå universitet, Umeå, 09:00 (Engelska)
Opponent
Handledare
Tillgänglig från: 2013-04-26 Skapad: 2013-04-24 Senast uppdaterad: 2018-06-08Bibliografiskt granskad

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Petzold, KatjaOlofsson, AnnelieArnqvist, AnnaGröbner, GerhardJürgen, Schleucher

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