Disulfide linkages reduction

A second kind of covalent bonding in peptides occurs when a disulfide linkage, RS-SR, is formed between two cysteine residues. As we saiv in Section 18.8, a disulfide is formed by mild oxidation of a thiol, RSH, and is cleaved by mild reduction. 

Fungal cutinases show no free SH groups but have 4 Cys residues, indicating that they are in disulfide linkage [119]. The reaction of the native enzyme with DTE was extremely slow but in the presence of SDS at its CMC rapid reduction could be observed [102]. Reduction of the disulfide bridge resulted in irreversible inactivation of the enzyme and the protein tended to become insoluble. CD spectra of cutinase in the 205-230 nm region, before and after DTE reduc-... 

Figure 5.3 SMPT can form crosslinks between an amine-containing molecule and a sulfhydryl-containing compound through amide and disulfide linkages, respectively. The hindered nature of the disulfide group provides better stability toward reduction and cleavage.

Figure 11.7 Sulfo-NHS-SS-biotin reacts with amine groups to form amide bonds. The biotin group can be later cleaved off the modified molecule by reduction of its internal disulfide linkage.

Figure 14.10 The crosslinker SPDP can be reacted with amine particles to create thiol-reactive pyridyl disulfide groups on the surface. Thiol-containing proteins or other thiol molecules can be reacted with these activated particles to result in disulfide linkages, which are reversible by reduction.

SPDP also is a popular choice for coupling sulfhydryl-containing molecules to liposomes. PE residues in vesicles may be activated with this crosslinker to form pyridyl disulfide derivatives that can react with sulfhydryls to form disulfide linkages. Unlike the iodoacetyl- and maleimide-based crosslinkers discussed previously, the linkage formed with SPDP is reversible by simple disulfide reduction. Pure PE may be activated with SPDP prior to its incorporation into a liposome,... 

It was a surprise to discover that a mutant of E. coli lacking thioredoxin can still reduce ribonucleotides. In the mutant cells thioredoxin is replaced by glutaredoxin, whose active site disulfide linkage is reduced by glutathione rather than directly by NADPH. Oxidized glutathione is, in turn, reduced by NADPH and glutathione reductase. Thus, the end result is the same with respect to ribonucleotide reduction. 

Applications of the TASP approach are then considered for such aspects as four-helical bundles (Section 13.1.2.4.2)J4 The details of structures and synthetic routes for the preparation of the bundles are described. Other chain assemblies and applications are discussed (Section 13.1.2.5) including metal-ion assisted self assemblies, disulfide linkages in the formation of four-helical bundles (Section 13.1.2.5.2), multiple antigenic peptide systems (Section 13.1.2.5.4), and templates constructed from a trialdehyde template by reductive amination to form a three-helical bundle (Section 13.1.2.5.5))5 ... 

Sodium tetrathionate (Na2S406) is a redox compound that under the right conditions can facilitate the formation of disulfide bonds from free sulfhydryls. The tetrathionate anion reacts with a sulfhydryl to create a somewhat stable active intermediate, a sulfenylthiosulfate (Fig. 102). Upon attack of the nucleophilic thiolate anion on this activated species, the thiosulfate (S203 =) leaving group is removed and a disulfide linkage forms (Pihl and Lange, 1962). The reduction of tetrathionate to thiosulfate in vivo was a subject of early study (Theis and Freeland, 1940 Chen et al., 1934). 

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