Lipid–Protein Interactions in Rat Renal Subcellular Membranes: A Biophysical and Biochemical Study

D’Antuono, Cecilia L.; Fernández-Tomé, Marı́a del Carmen; Sterín-Speziale, Norma; Bernik, Delia L.

doi:10.1006/abbi.2000.1979

Cited by 16 publications

(18 citation statements)

References 42 publications

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“…Asymmetric lipid distributions between the inner and outer bilayer leaflets (35)(36)(37) were further considered to specifically model the composition of the inner leaflet of the plasma membrane (Table 1). To simulate the composition of a lipid raft (38 -41), we mixed sphingomyelin and cholesterol with zwitterionic PE and anionic PS ( Table 1).…”

Section: Methodsmentioning

confidence: 99%

Lipid Rafts Alter the Stability and Activity of the Cholera Toxin A1 Subunit

Ray

Taylor

Banerjee

et al. 2012

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Lipid Rafts Alter the Stability and Activity of the Cholera Toxin A1 Subunit

Ray

Taylor

Banerjee

et al. 2012

Journal of Biological Chemistry

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“…Membrane lipids were extracted by the method of Bligh and Dyer [23] and resolved by silica gel thin-layer chromatography in accordance with the method described by D’Antuono et al [24]. SM, phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidylserine (PS) and phosphatidylethanolamine (PE) were well separated with Rf values of 0.18, 0.35, 0.45, 0.70 and 0.85, respectively.…”

Section: Methodsmentioning

confidence: 99%

Expression and Activity of SGLT2 in Diabetes Induced by Streptozotocin: Relationship with the Lipid Environment

Borghese

Majowicz²,

Ortiz³

et al. 2009

Nephron Physiol

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Background/Aims: Diabetes mellitus may impact on the regulation of renal Na⁺-glucose cotransporter type 2 (SGLT2), however, previous studies have yielded conflicting results on the effects of streptozotocin (STZ)-induced diabetes on SGLT-mediated glucose transport. Methods: Diabetes was induced in male Wistar rats. The studies were performed at 3 (D3), 7 (D7) and 14 (D14) days after a single i.p. injection of STZ. SGLT2 activity was measured using α-¹⁴C-methyl glucose uptake in brush-border vesicles (BBV) from renal cortex, and SGLT2 expression was assessed by immunoblotting. Phospholipids were quantified by a modification of Fiske-Subarow‘s method after being separated by thin-layer chromatography. Results: Glucose uptake was reduced in all groups of diabetic rats. SGLT2 expression decreased in D3 and D7. There was a decrease in sphingomyelin (SM) content and an increase in phosphatidylcholine (PC) content in BBV from D14 versus control, without differences in phosphatidylinositol (PI), phosphatidylserine (PS) and phosphatidylethanolamine (PE). Conclusion: The downregulation of SGLT2 activity during STZ-induced diabetes may be a protective mechanism to control the excess of circulating glucose and could be a consequence of a decrease in SGLT2 expression in D3 and D7, whereas altered activity of SGLT2 in D14 could be a consequence of changes in membrane lipid composition. However, we cannot discard the possibility that the decrease in SGLT2 activity could be due to a covalent modification of the active site of the protein.

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“…Many proteoliposome studies attempt to reproduce a typical membrane by mixing a few commercial lipids as a base composition [24–26], but, logically, using extracted endogenous membrane phospholipids would even more closely mimic the native lipid bilayer including lipid rafts. Functional protein reconstitution into vesicles is dependent on method of preparation and lipid composition of the vesicles, necessitating determination of degree of incorporation of a protein into the liposomes before detailed protein function can be reliably and repeatedly evaluated.…”

Section: Introductionmentioning

confidence: 99%

Lipid Bilayer Composition Affects Transmembrane Protein Orientation and Function

Hickey

Buhr

2011

Journal of Lipids

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Sperm membranes change in structure and composition upon ejaculation to undergo capacitation, a molecular transformation which enables spermatozoa to undergo the acrosome reaction and be capable of fertilization. Changes to the membrane environment including lipid composition, specifically lipid microdomains, may be responsible for enabling capacitation. To study the effect of lipid environment on proteins, liposomes were created using lipids extracted from bull sperm membranes, with or without a protein (Na+ K+-ATPase or α-amylase). Protein incorporation, function, and orientation were determined. Fluorescence resonance energy transfer (FRET) confirmed protein inclusion in the lipid bilayer, and protein function was confirmed using a colourometric assay of phosphate production from ATP cleavage. In the native lipid liposomes, ATPase was oriented with the β subunit facing the outer leaflet, while changing the lipid composition to 50% native lipids and 50% exogenous lipids significantly altered this orientation of Na+ K+-ATPase within the membranes.

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Lipid–Protein Interactions in Rat Renal Subcellular Membranes: A Biophysical and Biochemical Study

Cited by 16 publications

References 42 publications

Lipid Rafts Alter the Stability and Activity of the Cholera Toxin A1 Subunit

Lipid Rafts Alter the Stability and Activity of the Cholera Toxin A1 Subunit

Expression and Activity of SGLT2 in Diabetes Induced by Streptozotocin: Relationship with the Lipid Environment

Lipid Bilayer Composition Affects Transmembrane Protein Orientation and Function

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