Spin Labeling in Disease

Butterfield, D. Allan

doi:10.1007/978-1-4615-6540-6_1

Cited by 87 publications

(70 citation statements)

References 288 publications

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“…There is no difference in the WI S ratio of control and 50 jiM HNE-treated samples that were pretreated with GEE. lifetime of a particular orientation of the principal axis of the spin probe with respect to the static external magnetic field (Butterfield, 1982). In the EPR spectrum, this effect manifests as a line broadening, consequently leading to an increase in the HWHH (Butterfield, 1982;Hall et al, 1995a,b).…”

Section: Fig 5 Effect Ofpreincubation Of Synaptosomes With 2 MM Geementioning

confidence: 99%

“…One of the most widely used protein-specific spin labels to study membrane protein conformational changes is MAL-6 (Butterfield, 1982) (Fig. Ia) (Fig.…”

Section: Spin Labeling Cortical Synaptosomal Membrane Proteins With Mmentioning

confidence: 99%

“…The ratio of the EPR spectral amplitudes of the weakly immobilized line (W) to that of the strongly immobilized line (5), in the low-field region of the EPR spectrum of MAL-6-labeled synaptosomal membranes, is the W/S ratio. This parameter is known to be highly sensitive to protein conformational changes and protein-protein interactions (Butterfield, 1982;Trad and Butterfield, 1994;Bellary et al, 1995;Hall et al, 1995aHall et al, -c, 1997Howard et al, 1996;Butterfield et al, 1997b). Increased protein-protein interactions, decreased segmental motion, and/or conformational changes in the spin-labeled proteins lead to decreased molecular motion resulting in a lowering of the W/S ratio (Butterfield, 1982;Wyse and Butterfield, 1988;Trad and Butterfield, 1994;Bellary et al, 1995;Hall et al, 1995aHall et al, -c, 1997Howard et al, 1996;Butterfield et al, l997b).…”

Section: Spin Labeling Cortical Synaptosomal Membrane Proteins With Mmentioning

confidence: 99%

“…To obtain information about the synaptosomal membrane lipid dynamics and order, nitroxide stearate molecules are used as probes (Butterfield, 1982). Whereas MAL-6 binds to membrane proteins via a covalent bond, the nitroxide stearates intercalate in the bilayer with the charged head group near the head group of the phospholipids and the acyl chain of the probe aligned with the lipid acyl chains.…”

Section: Effect Of Hne Treatment On Motion and Order Of Phospholipidsmentioning

confidence: 99%

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The Lipid Peroxidation Product, 4‐Hydroxy‐2‐trans‐Nonenal, Alters the Conformation of Cortical Synaptosomal Membrane Proteins

Subramaniam

Roediger

Jordan³

et al. 1997

Journal of Neurochemistry

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254

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Alzheimer's disease (AD) is widely held to be a disorder associated with oxidative stress due, in part, to the membrane action of amyloid~3-peptide (A/3). A~-associated free radicals cause lipid peroxidation, a major product of which is 4-hydroxy-2-trans-nonenal (HNE). We determined whether HNE would alter the conformation of synaptosomal membrane proteins, which might be related to the known neurotoxicity of A~3and HNE. Electron paramagnetic resonance spectroscopy, using a protein-specific spin label, MAL-6 (2,2 ,6,6-tetramethyl-4-maleimidopiperidin-1-oxyl), was used to probe conformational changes in gerbil cortical synaptosomal membrane proteins, and a lipid-specific stearic acid label, 5-nitroxide stearate, was used to probe for HNE-induced alterations in the fluidity of the bilayer domain of these membranes. Synaptosomal membranes, incubated with low concentrations of HNE, exhibited changes in protein conformation and bilayer order and motion (fluidity). The changes in protein conformation were found to be concentration-and time-dependent. Significant protein conformational changes were observed at physiologically relevant concentrations of 1-10 p~MHNE, reminiscent of similar changes in synaptosomal membrane proteins from senile plaque-and A/3-rich AD hippocampal and inferior parietal brain regions. HNE-induced modifications in the physical state of gerbil synaptosomal membrane proteins were prevented completely by using excess glutathione ethyl ester, known to protect neurons from HNEcaused neurotoxicity. Membrane fluidity was found to increase at higher concentrations of HNE (50 /2M). The results obtained are discussed with relevance to the hypothesis of A/3-induced free radical-mediated lipid peroxidation, leading to subsequent HNE-induced alterations in the structure and function of key membrane proteins with consequent neurotoxicity in AD brain.

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Section: Fig 5 Effect Ofpreincubation Of Synaptosomes With 2 MM Geementioning

confidence: 99%

“…One of the most widely used protein-specific spin labels to study membrane protein conformational changes is MAL-6 (Butterfield, 1982) (Fig. Ia) (Fig.…”

Section: Spin Labeling Cortical Synaptosomal Membrane Proteins With Mmentioning

confidence: 99%

Section: Spin Labeling Cortical Synaptosomal Membrane Proteins With Mmentioning

confidence: 99%

Section: Effect Of Hne Treatment On Motion and Order Of Phospholipidsmentioning

confidence: 99%

See 2 more Smart Citations

The Lipid Peroxidation Product, 4‐Hydroxy‐2‐trans‐Nonenal, Alters the Conformation of Cortical Synaptosomal Membrane Proteins

Subramaniam

Roediger

Jordan³

et al. 1997

Journal of Neurochemistry

Self Cite

254

137

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“…In particular, the spin label Mal-6 was reported to react specifically with -SH groups of proteins, in particular spectrin, band 3, band 2.1, and other high molecular weight proteins (7). Spectra with two different correlation times for the motion were usually observed in erythrocyte ghost studies (8)(9)(10).…”

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confidence: 99%

Changes in Erythrocyte Properties during the First Hours of Life: Electron Spin Resonance of Reacting Sulfydryl Groups

et al. 1988

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ABSTRACT. In an attempt to clarify the mechanism by which the red cells (RBC) of newborn infants are protected against oxidative agents, electron spin resonance (ESR) assays were carried out using the nitroxide radical 4-maleimide-2,2,6,6-tetramethylpiperidinyl-1-01 (Mald), a sulfydryl-reacting agent. The ESR assays were performed in 24 samples of cord blood, 20 samples of blood from 4-day-old infants, and eight samples of &h-old infants. The analyses were carried out on whole blood and washed erythrocytes were resuspended in buffered saline. The same experiments were performed in 10 blood samples from healthy adults as controls. Whole blood, before and after removing the buffy coat, and cell-free plasma were also examined by ESR assay. Cell-free plasma and buffy coats proved not to be appreciably involved in the Ma14 behavior. The data of the ESR spectroscopy demonstrated a significantly slower reaction rate in the samples of cord blood and in blood of 8-h-old infants, compared to that of 4-day-old infants and adults. No significant differences in Mal-6 behavior could be detected between cord blood and 4-day-old infant blood in the results of ESR assays performed in washed red cells. Chemical determination of RBC-reacting sulfydryl groups and the assays of glutathione also demonstrated the absence of differences between cord blood and blood of 4-day-old infants. The results of our investigation suggest that the RBC-sulfydryl-reacting groups are less involved in the detoxification of oxidative agents during the first hours of life than in the following days. This peculiarity of RBC of younger infants appears to be due, to a considerable extent, to the modulation by plasma factors of the interactions between Mal-6 and RBCreacting sulfydryl groups. Therefore, the changes in plasma components occurring during the first hours of life appear to modify the interactions between the RBC and the oxidative agents. (Pediatr Res 24: 391-395, 1988) Abbreviations RBC, red blood cells ESR, electron spin resonance -SH, sulfydryl-reacting groups Mal-6,4-maleimide-2,2,6,6-tetramethylpiperidiny1-1-oxy1 Htc, hematocrit GSH, glutathione

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Site-directed and random immobilization of subtilisin on functionalized membranes: Activity determination in aqueous and organic media

Viswanath

Wang

Bachas

et al. 1998

Biotechnol. Bioeng.

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Kinetic comparisons have been made between a randomly immobilized and a site‐specifically immobilized subtilisin BPN′ on microfiltration membranes of varying hydrophilicities in both aqueous and organic media. Site‐directed mutagenesis was employed to introduce a single cysteine into the amino acid sequence of subtilisin at a location away from the active site. Immobilization of this mutant enzyme was then carried out using the single cysteine residue to orient the active site of the enzyme away from the membrane surface. Kinetic comparison of the immobilized mutant enzyme with the randomly immobilized wild‐type enzyme in aqueous media showed an activity enhancement on both hydrophilic silica‐containing and hydrophobic poly(ether)sulfone membranes. Higher loading efficiencies were observed for the site‐directed enzyme on immobilization. Optimal enzyme loading values were calculated for the randomly immobilized enzyme. An enhancement of activity was also observed for the site‐directed immobilized systems using nearly anhydrous hexane as the solvent. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 60: 608–616, 1998.

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Spin Labeling in Disease

Cited by 87 publications

References 288 publications

The Lipid Peroxidation Product, 4‐Hydroxy‐2‐trans‐Nonenal, Alters the Conformation of Cortical Synaptosomal Membrane Proteins

The Lipid Peroxidation Product, 4‐Hydroxy‐2‐trans‐Nonenal, Alters the Conformation of Cortical Synaptosomal Membrane Proteins

Changes in Erythrocyte Properties during the First Hours of Life: Electron Spin Resonance of Reacting Sulfydryl Groups

Site-directed and random immobilization of subtilisin on functionalized membranes: Activity determination in aqueous and organic media

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