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Effect of different surfactants and protocol changes (temperature and ionic strength changes, cholesterol depletion and local anesthetics) in detergent resistant membranes of erythrocytes

Grant number: 09/00904-1
Support Opportunities:Regular Research Grants
Duration: June 01, 2009 - November 30, 2011
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal Investigator:Eneida de Paula
Grantee:Eneida de Paula
Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil


Lateral phase separation forms membrane domains (lipid rafts) of different composition required for specific proteins to maintain their function. This biomembranes' phenomenon has been associated to process such as fusion, intercellular communication, signal transduction and nervous impulse propagation. Erythrocyte membrane is a really good model for lipid rafts studies. The properties of these domains have been, operationally studied through the use of cholesterol/sphingolipids enriched fractions, called DRMs (detergent-resistant membranes), obtained with surfactants, at low temperature. Our research group (Biomembranes, IB/Unicamp) in collaboration with Dr. Giampaolo Minetti's group (Universita Degli Studi di Pavia, Itália) has recently demonstrated erythrocytes' DRMs formation with the use of the non ionic detergent C12E8 (Domingues et al, 2009). With the present financial request we intend to acquire the necessary equipments and reagents in order to continue with this line of investigation, in an independent way. We intend to evaluate: i) the effect of other non ionic detergents (Brij, Tween,...) to induce DRM formation and ii) changes in the protocol of DRM preparation such as temperature, cholesterol depletion and presence of local anesthetics in the lipidic and proteic composition of these rafts. Our main goal is to get information on the molecular interaction between detergents and membrane components, in order to relate them to the biomembranes organization and function, with emphasis in the solubilization process. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
CASADEI, BRUNA RENATA; CARVALHO, PATRICIA DE OLIVEIRA; RISKE, KARIN A.; BARBOSA, RAQUEL DE MELO; DE PAULA, ENEIDA; DOMINGUES, CLEYTON CREPALDI. Brij detergents reveal new aspects of membrane microdomain in erythrocytes. MOLECULAR MEMBRANE BIOLOGY, v. 31, n. 6, p. 195-205, . (10/18516-5, 09/00904-1)
DOMINGUES, CLEYTON C.; CIANA, ANNARITA; BUTTAFAVA, ARMANDO; CASADEI, BRUNA RENATA; BALDUINI, CESARE; DE PAULA, ENEIDA; MINETTI, GIAMPAOLO. Effect of Cholesterol Depletion and Temperature on the Isolation of Detergent-Resistant Membranes from Human Erythrocytes. Journal of Membrane Biology, v. 234, n. 3, p. 195-205, . (09/00904-1)
CLOP, EDUARDO M.; FRACETO, LEONARDO F.; MIGUEL, VIRGINIA; GASTALDI, SALOME; DE PAULA, ENEIDA; PERILLO, MARIA ANGELICA. Combined in-silico and in-vitro experiments support acid-base equilibrium as a tool to estimate the localization depth of 4-nitrophenol within a phospholipid bilayer. BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, v. 1864, n. 11, p. 11-pg., . (09/00904-1)

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