The development of novel thiol reagents for probing ion channel structure

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National Library of Canada , Ottawa
SeriesCanadian theses = -- Thèses canadiennes
The Physical Object
FormatMicroform
Pagination1 microfiche : negative. --
ID Numbers
Open LibraryOL19364190M
ISBN 100612454711
OCLC/WorldCa51536449

We have synthesized a novel thiol reagent, 2-[(methylsulfonyl)thio]ethyl [N-(N,N-dimethylamino)ethyl]carbamate (MTSAC), that contains a carbamate functional group as well as a (positively charged) terminal amino group. The carbamate C−N bond isomerizes on a millisecond time scale and significantly alters the three-dimensional shape of the by:   We have synthesized a novel thiol reagent, 2-[(methylsulfonyl) thio] ethyl [N-(N,N-dimethylamino) ethyl] carbamate (MTSAC), that contains a carbamate functional group as well as a (positively charged) terminal amino group.

The carbamate C-N bond isomerizes on a millisecond time scale and significantly alters the three-dimensional shape of the reagent. The behavior of this reagent Cited by: A. The Glutathione-Operated Cation Channel of Vascular Endothelial Cells.

The effects of sulfhydryl reagents on ion flux have been described for many types of ion channels (Chiamvimonvat et al., ; Koivisto et al., ; Lei et al., ). In large part, these studies have documented changes in either the magnitude of current (ionic. The thiol-reagent N-(6-(biotininamido)hexyl)-3′-(2′-pyridylthio)propionamide (biotin-HPDP) contains a biotin moiety and attaches The development of novel thiol reagents for probing ion channel structure book free thiols by means of a disulfide linkage.

It can therefore be used to specifically enrich for the oxidized protein-thiol fraction of the proteome (Jaffrey and Snyder, ; Le Moan et al., ).

from book Novel Chemical Tools to Study Ion mutagenesis to probe structure-function relationships in ion channels focusing mainly on Cys-loop receptors. inhibited not only by thiol. Development of a Novel Thiol Reagent for Probing Ion Channel Structure: Studies in a Model System.

Biochemistry36 (6), DOI: /bi   This review has focused on three aspects of ion-channel engineering, with an emphasis on voltage-gated channels: (1) modifying ion-channel parts to understand how they function, (2) tailoring ion channels to probe cell biology and physiology, and (3) engineering novel modulators of ion channels.

The development of novel thiol reagents for probing ion channel structure: Foong, Louise Yuli: Thiol reactive chemical probes for studying protein ion channel structure and the application of chemical probes to the study of U2AF, an essential RNA splicing factor.

Polar Reagents for Determining Thiol Accessibility; 1,Phenanthroline Iodoacetamide for Preparing Metal-Binding Conjugates; Related Tables. Thiol-reactive dyes excited with visible light—Table ; Thiol-reactive dyes excited with ultraviolet light—Table Akabas M.

() Cysteine Modification: Probing Channel Structure, Function and Conformational Change. In: Ahern C., Pless S. (eds) Novel Chemical Tools to Study Ion Channel Biology. Advances in Experimental Medicine and Biology, vol   a Strategy for probing free cysteine thiols that become exposed to the solvent upon protein unfolding and permissive to maleimide reaction.

b Structure of tetraphenylethene (TPE) conjugated to a. Abstract. Chloride ion channels have been found to play crucial roles in the development of human diseases, for example, mutations in the genes encoding Cl − channels lead to a variety of deleterious diseases in muscle, kidney, bone, and brain, including myotonia congenita, dystrophia myotonica, cystic fibrosis, osteopetrosis, and epilepsy, and similarly their activation is supposed to be.

Upon thiol addition, the conjugation is disrupted and the fluorescence of the coumarin fluorophore is restored. The probe 38 is a highly sensitive thiol reagent showing over fold increases in fluorescence (λ ex nm, λ em nm) by forming 39 through Michael addition.

The detection limit was found to be 1 μM for Cys in 25 mM phosphate. This section describes a variety of probes for Ca 2+, Na +, K + and Cl – ion channels and carriers.

Indicators for Ca2+, Mg2+, Zn2+ and Other Metal Ions—Chap pH Indicators—Chapter 20 and Indicators for Na+, K+, Cl– and Miscellaneous Ions—Chapter 21 contain our extensive selection of indicators for these physiologically important ions, providing a means of correlating ion.

porter ion quantification. HeLa proteomes were labeled with the alkyne-tagged thiol-reactive re-agent, IPM, and digested into tryptic peptides.

The digest mixture was split into eight identical aliquots. Each was then labeled with one of the eight isobaric iTRAQ reagents, and the deriv-atized digests combined in a predefined ratio ( Most of Molecular Probes’ thiol-reactive reagents will react with thiol groups on proteins to give thioether-coupled products.

These reagents react rapidly at near-neutral (physiological) pH and usually can be coupled with thiol groups selectively in the presence of amine groups.1,2.

Details The development of novel thiol reagents for probing ion channel structure FB2

However, probing of the intact channel requires access of the thiol reagent to the cytoplasmic opening in an intact cell pair through an environment that is highly reducing.

Passive perfusion through whole cell patch clamps was inadequate for this purpose, which required active perfusion of the cytoplasm. This data can complement the structural insights obtained from the burgeoning number of crystal structures of detergent solubilized membrane proteins whose functional state is often uncertain.

This article will review the use of cysteine mutagenesis to probe structure-function relationships in ion channels focusing mainly on Cysloop receptors. KcsA, a potassium channel from Streptomyces lividans, is the ion channel that is best understood structurally.

The structure of KcsA has been solved in. A biochemically unique cone snail venom peptide has been characterized that may be used to probe unexplored but important features of the diverse voltage-gated Na channel isoforms that underlie electrical signaling in the nervous system.

This peptide has a unique posttranslational modification (S-cysteinylated cysteine) and blocks sodium channels by forming a disulfide bond with the channel at. Thiols are important molecules in the environment and in biological processes.

Cysteine (Cys), homocysteine (Hcy), glutathione (GSH) and hydrogen sulfide (H2S) play critical roles in a variety of physiological and pathological processes.

The selective detection of thiols using reaction-based probes and sensors is very important in basic research and in disease diagnosis.

Description The development of novel thiol reagents for probing ion channel structure EPUB

Toxins have evolved to target regions of membrane ion channels that underlie ligand binding, gating, or ion permeation, and have thus served as invaluable tools for probing channel structure and.

3 Thiol and Sulfide Quantitation Kit Add mL of Buffer B (see Reagents Required but Not Provided) to each of two clean tubes. Add mL of the mM L-cysteine working solution to one of the tubes and mL of Buffer A to the other, to serve as a.

Optimized thiol derivatizing reagent for the mass spectral analysis of disubstituted epoxy fatty acids John W.

Newman, Bruce D. Hammock* Department of Entomology and the University of California Davis Cancer Research Center,University of California, Briggs Hall, 1Shields Avenue,Davis,CA ,USA. In the 90s, the development of a novel single molecule technique based on nanopore sensing emerged.

Preliminary improvements were based on the molecular or biological engineering of protein nanopores along with the use of nanotechnologies developed in the context of microelectronics.

Since the last decade, the convergence between those two worlds has allowed for biomimetic approaches. © Alison Frontier, University of Rochester.

Supported by a grant from the National Science Foundation. NSF Funding {+} This material is based upon work supported by the National Science Foundation under Grant Number CHE Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect.

Sigma-Aldrich Online Catalog Product List: Thiols. Using the channel-permeant thiol probe Au[CN] 2 −, which has a size similar to chloride (Smith et al., ), Gao and Hwang () showed that IC, FC, and TC (residues external to the narrow region) are only reactive to internal Au[CN] 2 − in the presence of ATP but not in the absence of ATP (see Fig.

7 B for relative position. The ATP-powered AAA+ protease, ClpXP, degrades the thiol oxidative transcription regulator, Spx, under non-stress conditions. The novel protein YjbH facilitates ClpXP recognition of the Spx C-terminal, possibly by interacting with the C-terminal part of Spx and thereby making it more accessible, resulting in rapid degradation of Spx by the.

Chapter 2.

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Thiol-Methacrylate Networks and Its Application in Novel Hydrogels (Claudia I. Vallo and Silvana V. Asmussen, Institute of Materials Science and Technology, University of Mar del Plata, CONICET, Mar del Plata, Argentina) Chapter 3.

Thionyl Chloride (SOCl2)-Mediated Homocoupling of Thiols into Disulfides at Ambient Conditions. Mutations in cystic fibrosis transmembrane conductance regulator (CFTR) cause cystic fibrosis, a lethal genetic disease occurring in people of northern European descent.

Decades of study have been directed toward a molecular understanding of this ion channel. The structure presented here enables a direct correlation of structure with function, most of which has been characterized in .embryo during development. Confocal microscopy of the embryo, followed by in-gel staining and imaging of proteins, then tandem MS, led to our discovery of two novel disulfide-containing proteins in the fish chorion, lipovitellin and C-reactive protein (CRP).

CRP is a well-known biomarker for infection or inflammation (associated with.A compound represented by the following formula (I), which is useful as a reagent with which the concentrations of a peptide and a protein can be efficiently and easily determined with high sensitivity; and a method of analysis in which a reagent comprising the compound is used.

[Chemical formula 1] Formula (I) [In the formula, R1 represents optionally substituted aryl or heteroaryl; R3, R4.