Coupling thiol groups

Thiolated chitosans are synthesized by covalently coupling thiol groups to chitosan backbones. They exhibit in situ gel-forming properties owing to the formation of... 

The synthesis of polysulfide elastomers involves the use of a small amount of trichloroalkane in addition to dichloroalkane and sodium sulfide in order to form a branched polymer. The prepolymer is treated with a mixture of sodium hydrosulfide and sodium sulfite followed by acidification to convert all end-groups to thiol groups. Further polymerization and crosslinking is achieved by oxidative coupling of the thiol end-groups by treatment with lead dioxide, p-quinone dioxime, or other oxidizing agent... 

Figure 7.26 Dendrimers made with a disulfide-containing core can be reduced to produce dendrons having free thiol groups for surface modification. Dative binding of these thiol-dendrons to gold or metallic surfaces can provide a high density of amine groups for coupling proteins or other molecules.

Figure 9.34 The long side chain of this BODIPY derivative contains a sulfhydryl-reactive iodoacetamide group that can couple to a thiol group to form a thioether bond.

Add 1-10 mg of a protein or antibody containing an available thiol group to the particle suspension. Alternatively, add the protein to be coupled to the particle suspension in an amount equal to 1-10 X molar excess over the calculated monolayer for the protein type to be coupled. The optimal amount of protein to be added should be determined experimentally. Creating thiol groups on proteins or peptides may be done from disulfides by reduction. Alternatively, a thiolation reagent may be used to add thiols to the protein surface for coupling (see the protocols in Chapter 1, Section 4.1). 

Add 1-10 mg of a protein or antibody containing an available thiol group to the particle suspension in coupling buffer and mix to dissolve. Alternatively, add the protein to the particle suspension in an amount equal to 1-10 X molar excess over the calculated... 

Figure 21.11 Fab antibody fragments containing free thiols can be activated with Ellman s reagent to form a sulfhydryl-reactive derivative. A-chain toxin subunits containing a free thiol group may be coupled to the activated Fab molecule to produce an immunotoxin complex.

Many of the chemical derivatization methods employed in these strategies involve the use of an activation step that produces a reactive intermediary. The activated species then can be used to couple a molecule containing a nucleophile, such as a primary amine or a thiol group. The following sections describe the chemical modification methods suitable for derivatizing individual nucleic acids as well as oligonucleotide polymers. 

Reduction of the cystamine-labeled oligo using a disulfide reducing agent releases 2-mer-captoethylamine and creates a thiol group for conjugation (Figure 27.6). DNA probes labeled in this manner have been successfully coupled with SPDP-activated alkaline phosphatase (Chapter 26, Sections 1.2 and 2.5), maleimide-activated horseradish peroxidase (HRP) (Chapter 26, Section 1.1), NHS-LC-biotin (Chapter 11, Section 1 and Chapter 27, Section 2.3), and the fluorescent tag AMCA-HPDP (Chapter 9, Section 3 and Chapter 27, Section 2.5). 

Figure 28.18 FeBABE can be coupled to an available thiol group on a bait protein using the bromoacetyl reactive group to form a thioether linkage.

Moroney, J.V., Wamcke, K., and McCarthy, R.F. (1982) The distance between thiol groups in the gamma subunit of coupling factor 1 influences the protein permeability of thylakoid membranes. J. Bioenerg. Biomembr. 14, 347. 

Fig. 19. Lipopeptides with the aminoacid sequence of the Ras C-terminus and the natural or artificial lipid-modifications can be coupled with C-terminally truncated Ras via a maleimi-docaproyl linker. This electrophile reacts with free thiol groups (here, a C-terminal cysteine at the Ras moiety)...

The microspheres—synthesised via a two-step process (acid-catalysed hydrolysis and condensation of 3-mercaptopropyltrimethoxysilane (MPS) in aqueous solution, followed by condensation catalysed by triethanolamine)—have a narrow size distribution (Figure 5.16) and are considerably more stable than polystyrene divinylbenzene microspheres as shown in phosphoramidite oligonucleotide synthesis by the excellent retention of fluorescence intensity in each of the reagent steps involved in phosphoramidite DNA synthesis (Figure 5.17, in which the organo-silica microsphere free thiol groups are derivatized with ATTO 550 maleimide coupled to the entrapped dye). 

Tsarevsky, N. V. Matyjaszewski, K. Reversible redox cleavage/coupling of polystyrene with disulfide or thiol groups prepared by atom transfer radical polymerizaion. Macromolecules 2002, 35, 9009. 

Acyl residues are usually activated by transfer to coenzyme A (2). In coenzyme A (see p. 12), pantetheine is linked to 3 -phos-pho-ADP by a phosphoric acid anhydride bond. Pantetheine consists of three components connected by amide bonds—pantoic acid, alanine, and cysteamine. The latter two components are biogenic amines formed by the decarboxylation of aspartate and cysteine, respectively. The compound formed from pantoic acid and p-alanine (pantothenic acid) has vitamin-like characteristics for humans (see p. 368). Reactions between the thiol group of the cysteamine residue and carboxylic acids give rise to thioesters, such as acetyl CoA. This reaction is strongly endergonic, and it is therefore coupled to exergonic processes. Thioesters represent the activated form of carboxylic adds, because acyl residues of this type have a high chemical potential and are easily transferred to other molecules. This property is often exploited in metabolism. 

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