Perturbational actions of barbiturate analogues on erythrocyte and synaptosomal membranes

1985 ◽  
Vol 63 (8) ◽  
pp. 937-943 ◽  
Author(s):  
David V. Godin ◽  
Peter McGinn
Keyword(s):  
Membrane Proteins ◽  
Local Anaesthetics ◽  
Erythrocyte Membranes ◽  
Synaptosomal Membranes ◽  
Structural Components ◽  
Chemical Probes ◽  
Trinitrobenzenesulfonic Acid ◽  
Specific Chemical ◽  
Nitrobenzoic Acid ◽  
Acid Incorporation

The membrane perturbational actions of pentobarbital, methohexital, and phenobarbital were analyzed in terms of their effects on the incorporation of group-specific chemical probes into protein and phospholipid structural components and on the activity of various functionally distinct ATPases in erythrocyte and brain synaptosomal membranes. When compared at concentrations producing similar degrees of antihemolysis in red cells, the three analogues differed most markedly in the nature and degree of the structural and functional alterations induced in membrane proteins, with the most highly lipophilic agent, methohexital, being frequently, although not invariably, the most potent. Comparison of the effects of pentobarbital on characteristics of trinitrobenzenesulfonic acid and 5,5′-dithiobis-(2-nitrobenzoic acid) incorporation into erythrocyte membranes with results previously obtained using other anaesthetics at concentrations producing 50% antihemolysis showed that pentobarbital behaves similarly to neutral general anaesthetics but differently from cationic local anaesthetics. Our findings suggest that the membrane perturbational characteristics of barbiturates may contribute to their diverse and complex actions on excitable tissues.

Studies on the structure and function of 16S ribosomal RNA using structure-specific chemical probes

1985 ◽  
Vol 8 (3-4) ◽  
pp. 747-755 ◽  
Author(s):  
Harry F. Noller ◽  
Barbara J. Van Stolk ◽  
Danesh Moazed ◽  
Stephen Douthwaite ◽  
Robin R. Gutell
Keyword(s):  
Ribosomal Rna ◽  
Structure And Function ◽  
16S Ribosomal Rna ◽  
Chemical Probes ◽  
Specific Chemical ◽  
And Function

scholarly journals Proteome-Wide Survey Reveals Norbornene Is Complementary to Dimedone and Related Nucleophilic Probes for Cysteine Sulfenic Acid

2019 ◽  
Author(s):  
Lisa Alcock ◽  
Maike Langini ◽  
Kai Stühler ◽  
Marc Remke ◽  
Michael Perkins ◽  
...  
Keyword(s):  
Redox Regulation ◽  
Chemical Reactivity ◽  
Live Cells ◽  
Chemical Probes ◽  
Cysteine Oxidation ◽  
Sulfenic Acid ◽  
Proteomics Analysis ◽  
New Members ◽  
Specific Chemical ◽  
Future Studies

<p>Detection of cysteine sulfenic acid in live cells is critical in advancing our understanding of cysteine redox chemistry and its biological function. Accordingly, there is a need to develop sulfenic acid-specific chemical probes with distinct reaction mechanisms to facilitate proteome-wide detection of this important posttranslational modification. Herein, we report the first whole-cell proteomics analysis using a norbornene probe to detect cysteine sulfenic acid in live HeLa cells. Comparison of the enriched proteins to those identified using dimedone and other <i>C</i>-nucleophilic probes revealed a complementary reactivity profile. Remarkably, 148 new members of the sulfenome were identified. These discoveries highlight how subtle differences in chemical reactivity of both the probes and cysteine residues influence detection. Overall, this study expands our understanding of protein oxidation at cysteine and reveals new proteins to consider for future studies of cysteine oxidation, redox regulation and signaling, and the biochemistry of oxidative stress. </p>

scholarly journals Proteome-Wide Survey Reveals Norbornene Is Complementary to Dimedone and Related Nucleophilic Probes for Cysteine Sulfenic Acid

2019 ◽  
Author(s):  
Lisa Alcock ◽  
Maike Langini ◽  
Kai Stühler ◽  
Marc Remke ◽  
Michael Perkins ◽  
...  
Keyword(s):  
Redox Regulation ◽  
Chemical Reactivity ◽  
Live Cells ◽  
Chemical Probes ◽  
Cysteine Oxidation ◽  
Sulfenic Acid ◽  
Proteomics Analysis ◽  
New Members ◽  
Specific Chemical ◽  
Future Studies

<p>Detection of cysteine sulfenic acid in live cells is critical in advancing our understanding of cysteine redox chemistry and its biological function. Accordingly, there is a need to develop sulfenic acid-specific chemical probes with distinct reaction mechanisms to facilitate proteome-wide detection of this important posttranslational modification. Herein, we report the first whole-cell proteomics analysis using a norbornene probe to detect cysteine sulfenic acid in live HeLa cells. Comparison of the enriched proteins to those identified using dimedone and other <i>C</i>-nucleophilic probes revealed a complementary reactivity profile. Remarkably, 148 new members of the sulfenome were identified. These discoveries highlight how subtle differences in chemical reactivity of both the probes and cysteine residues influence detection. Overall, this study expands our understanding of protein oxidation at cysteine and reveals new proteins to consider for future studies of cysteine oxidation, redox regulation and signaling, and the biochemistry of oxidative stress. </p>

scholarly journals Lysosome Targeting Chimeras (LYTACs) That Engage a Liver-Specific Asialoglycoprotein Receptor for Targeted Protein Degradation

2020 ◽  
Author(s):  
Green Ahn ◽  
Steven Banik ◽  
Caitlyn L. Miller ◽  
Nicholas Riley ◽  
Jennifer R. Cochran ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Protein Degradation ◽  
Pharmacokinetic Profile ◽  
Asialoglycoprotein Receptor ◽  
Surface Proteins ◽  
New Paradigm ◽  
Specific Chemical ◽  
Chemical Conjugation ◽  
Reduce Cell Proliferation

<p>Selective protein degradation platforms have afforded new development opportunities for therapeutics and tools for biological inquiry. The first lysosome targeting chimeras (LYTACs) targeted extracellular and membrane proteins for degradation by bridging a target protein to an endogenous lysosome targeting receptor, the cation-independent mannose-6-phosphate receptor (CI-M6PR). Here we developed LYTACs that engage the asialoglycoprotein receptor (ASGPR), a liver-specific lysosomal targeting receptor, to degrade membrane proteins in a tissue-specific manner. We conjugated antibody-based binders targeting cell-surface proteins to a tri-GalNAc motif that engages ASGPR. The resulting LYTACs directed lysosome trafficking of the bound targets and subsequent degradation. Degradation mediated by an EGFR-targeted GalNAc-LYTAC resulted in significant functional effects on the downstream kinase signaling of EGFR compared to canonical inhibition with a monoclonal antibody. Furthermore, we demonstrated that a small target binder, a 3.4 kDa peptide, can be linked to a single tri-GalNAc ligand to degrade integrins and significantly reduce cell proliferation. Site-specific chemical conjugation of one or two tri-GalNAc ligands to antibody scaffolds improved the pharmacokinetic profile of GalNAc-LYTACs <i>in vivo</i> compared to non-specific chemical conjugation. GalNAc-LYTACs represent an exciting new paradigm for cell-type restricted degradation of proteins.</p>

Interaction Mechanism of Cortisol and Catecholamines with Structural Components of Erythrocyte Membranes

2010 ◽  
Vol 114 (29) ◽  
pp. 9462-9473 ◽  
Author(s):  
Lev Evgenjevich Panin ◽  
Pavel Valentinovich Mokrushnikov ◽  
Valery Georgievich Kunitsyn ◽  
Boris Nikolaevich Zaitsev
Keyword(s):  
Interaction Mechanism ◽  
Erythrocyte Membranes ◽  
Structural Components

Chemical Probes for the Geometry of Membrane Proteins

1975 ◽  
pp. 19-38
Author(s):  
Frederic M. Richards ◽  
James V. Staros ◽  
Kuan Wang ◽  
H. Heitzmann
Keyword(s):  
Membrane Proteins ◽  
Chemical Probes

scholarly journals Novel pathogenic XK mutations in McLeod syndrome and interaction between XK protein and chorein

2019 ◽  
Vol 5 (3) ◽  
pp. e328 ◽  
Author(s):  
Yuka Urata ◽  
Masayuki Nakamura ◽  
Natsuki Sasaki ◽  
Nari Shiokawa ◽  
Yoshiaki Nishida ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Erythrocyte Membrane ◽  
Molecular Analysis ◽  
Cultured Cells ◽  
Immunoblot Analysis ◽  
Erythrocyte Membranes ◽  
Onset Age ◽  
Novel Mutations ◽  
Erythrocyte Membrane Proteins ◽  
Normal Controls

ObjectiveTo identify XK pathologic mutations in 6 patients with suspected McLeod syndrome (MLS) and a possible interaction between the chorea-acanthocytosis (ChAc)- and MLS-responsible proteins: chorein and XK protein.MethodsErythrocyte membrane proteins from patients with suspected MLS and patients with ChAc, ChAc mutant carriers, and normal controls were analyzed by XK and chorein immunoblotting. We performed mutation analysis and XK immunoblotting to molecularly diagnose the patients with suspected MLS. Lysates of cultured cells were co-immunoprecipitated with anti-XK and anti-chorein antibodies.ResultsAll suspected MLS cases were molecularly diagnosed with MLS, and novel mutations were identified. The average onset age was 46.8 ± 8 years, which was older than that of the patients with ChAc. The immunoblot analysis revealed remarkably reduced chorein immunoreactivity in all patients with MLS. The immunoprecipitation analysis indicated a direct or indirect chorein-XK interaction.ConclusionsIn this study, XK pathogenic mutations were identified in all 6 MLS cases, including novel mutations. Chorein immunoreactions were significantly reduced in MLS erythrocyte membranes. In addition, we demonstrated a possible interaction between the chorein and XK protein via molecular analysis. The reduction in chorein expression is similar to that between Kell antigens and XK protein, although the chorein-XK interaction is a possibly noncovalent binding unlike the covalent Kell-XK complex. Our results suggest that reduced chorein levels following lack of XK protein are possibly associated with molecular pathogenesis in MLS.

Effects of a series of alcohols on the binding of a fluorescent dye to erythrocyte membranes

1976 ◽  
Vol 54 (1) ◽  
pp. 35-41 ◽  
Author(s):  
S. H. Roth ◽  
L. Spero
Keyword(s):  
Membrane Proteins ◽  
Binding Sites ◽  
Erythrocyte Membranes ◽  
Membrane Action ◽  
Apparent Dissociation Constant ◽  
Ans Binding ◽  
Membrane Interaction ◽  
Low Concentrations ◽  
Anesthetic Concentration ◽  
Hypotonic Hemolysis

1. The effects of a series of aliphatic alcohols (methanol to octanol) on membrane proteins of erythrocytes were studied by monitoring the fluorescence of a dye (1-anilino-8-naphthalenesulfonic acid (ANS)) that adsorbs to erythrocyte ghost membranes. Low concentrations of all the alcohols reduced the ANS fluorescence of the membrane–ANS suspensions; lent to those which protect against hypotonic hemolysis on intact erythrocytes; higher concentrations markedly increased the fluorescence. Ethanol and methanol decreased ANS fluorescence at all concentrations.2. Lytic concentrations of saponin did not increase ANS fluorescence and did not modify the membrane action of the alcohols.3. None of these effects were observed in liposomes prepared from lipid extracts of the erythrocyte membrane.4. Since the apparent dissociation constant for the ANS–membrane interaction was unchanged in the presence of the alcohols, it was assumed that the fluorescence changes reflected alterations in the number of ANS binding sites. One conclusion is that low anesthetic concentration of the alcohols alter the conformation of membrane proteins, as indicated by the decreased number of ANS binding sites.

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