Dataset Information

The solubilisation of boar sperm membranes by different detergents - a microscopic, MALDI-TOF MS, (31)P NMR and PAGE study on membrane lysis, extraction efficiency, lipid and protein composition.

ABSTRACT: BACKGROUND: Detergents are often used to isolate proteins, lipids as well as "detergent-resistant membrane domains" (DRMs) from cells. Different detergents affect different membrane structures according to their physico-chemical properties. However, the effects of different detergents on membrane lysis of boar spermatozoa and the lipid composition of DRMs prepared from the affected sperm membranes have not been investigated so far. RESULTS: Spermatozoa were treated with the selected detergents Pluronic F-127, sodium cholate, CHAPS, Tween 20, Triton X-100 and Brij 96V. Different patterns of membrane disintegration were observed by light and electron microscopy. In accordance with microscopic data, different amounts of lipids and proteins were released from the cells by the different detergents. The biochemical methods to assay the phosphorus and cholesterol contents as well as 31P NMR to determine the phospholipids were not influenced by the presence of detergents since comparable amounts of lipids were detected in the organic extracts from whole cell suspensions after exposure to each detergent. However, matrix-assisted laser desorption and ionization time-of-flight mass spectrometry applied to identify phospholipids was essentially disturbed by the presence of detergents which exerted particular suppression effects on signal intensities. After separation of the membrane fractions released by detergents on a sucrose gradient only Triton X-100 and sodium cholate produced sharp turbid DRM bands. Only membrane solubilisation by Triton X-100 leads to an enrichment of cholesterol, sphingomyelin, phosphatidylinositol and phosphatidylethanolamine in a visible DRM band accompanied by a selective accumulation of proteins. CONCLUSION: The boar sperm membranes are solubilised to a different extent by the used detergents. Particularly, the very unique DRMs isolated after Triton X-100 exposure are interesting candidates for further studies regarding the architecture of sperm.

SUBMITTER: Jakop U

PROVIDER: S-EPMC2781011 | biostudies-literature | 2009

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

The solubilisation of boar sperm membranes by different detergents - a microscopic, MALDI-TOF MS, (31)P NMR and PAGE study on membrane lysis, extraction efficiency, lipid and protein composition.

Jakop Ulrike U Fuchs Beate B Süss Rosmarie R Wibbelt Gudrun G Braun Beate B Müller Karin K Schiller Jürgen J

Lipids in health and disease 20091111

<h4>Background</h4>Detergents are often used to isolate proteins, lipids as well as "detergent-resistant membrane domains" (DRMs) from cells. Different detergents affect different membrane structures according to their physico-chemical properties. However, the effects of different detergents on membrane lysis of boar spermatozoa and the lipid composition of DRMs prepared from the affected sperm membranes have not been investigated so far.<h4>Results</h4>Spermatozoa were treated with the selected ...[more]

PMID: 19906304

Similar Datasets

Project description:MALDI-TOF mass spectrometry (MS) on whole cells enables the detection of different cell types and cell activation. Here, we wondered whether this approach would be useful to investigate the host response in sepsis.Peripheral blood mononuclear cells (PBMCs) from patients with severe sepsis and healthy donors were analyzed with MALDI-TOF MS. PBMCs from healthy donors were also stimulated with lipopolysaccharide, peptidoglycan, CpG oligonucleotides, polyinosinic polycytidylic acid, and with heat-inactivated bacteria. Averaged spectra of PBMCs stimulated in vitro by different agonists were generated from the database using the Biotyper software and matching scores between each spectrum from patients and averaged spectra from the database were calculated.We show that the MALDI-TOF MS signature of PBMCs from septic patients was specific, compared with healthy controls. As the fingerprints observed in patients may be related to PBMC activation, PBMCs from healthy controls were stimulated with cytokines, soluble Pathogen-Associated Molecular Patterns (PAMPs) and heat-killed bacteria, and we created a database of reference spectra. The MALDI-TOF MS profiles of PBMCs from septic patients were then compared with the database. No differences were found between patients with documented infection (n?=?6) and those without bacteriological documentation (n?=?6). The spectra of PBMCs from septic patients matched with those of interferon-?- and interleukin-10-stimulated PBMCs, confirming that sepsis is characterized by both inflammatory and immunoregulatory features. Interestingly, the spectra of PBMCs from septic patients without documented infection matched with the reference spectrum of PBMCs stimulated by CpG oligonucleotides, suggesting a bacterial etiology in these patients.Despite the limits of this preliminary study, these results indicate that the monitoring of functional status of PBMCs in peripheral blood by whole cell MALDI-TOF MS could provide unique opportunities to assess disease progression or resolution in clinical settings.

Dataset Information

The solubilisation of boar sperm membranes by different detergents - a microscopic, MALDI-TOF MS, (31)P NMR and PAGE study on membrane lysis, extraction efficiency, lipid and protein composition.

Publications

The solubilisation of boar sperm membranes by different detergents - a microscopic, MALDI-TOF MS, (31)P NMR and PAGE study on membrane lysis, extraction efficiency, lipid and protein composition.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets