Modification cystamine

For reduction of the cystamine disulfides, add 20 pi of 1.0 M DTT and incubate at room temperature for 15 minutes. This will release 2-mercaptoethylamine from the cystamine modification site and create the free sulfhydryl on the 5 terminus of the oligonucleotide. 

The following protocol is useful for the modification of proteins with cystamine with subsequent reduction to create the free sulfhydryl. 

Modification of Nucleic Acids and Oligonucleotides with Cystamine... 

Figure 21.10 Cystamine may be used to make immunotoxin conjugates by a disulfide interchange reaction. Modification of antibody molecules using an EDC-mediated reaction creates a sulfhydryl-reactive derivative. A-chain toxin subunits containing a free thiol can be coupled to the cystamine-modified antibody to form disulfide crosslinks.

The following protocol describes the modification of DNA or RNA probes at their 5 -phosphate ends with a bis-hydrazide compound, such as adipic acid dihydrazide or carbohydrazide. A similar procedure for coupling the diamine compound cystamine can be found in Section 2.2 (this chapter). 

More recently, thiolated dextran was synthesised by modification of dextran with 4-nitrophenyl chloroformate in DMSO/pyridine in the presence of DMAP at 0 °C yielding a carbonate (content 6%), which can be substituted with cystamine. Subsequent reduction yields the thiolated dextran containing between 1 and 4% thiol moieties (Fig. 57) [370]. 

Regulation of liver D-fructose 1,6-diphosphatase could involve modification of the sulfhydryl group of the enzyme.397 Cystamine (2,2 -dithiobisethylamine) or homocysteine undergoes a disulfide exchange-reaction with two reactive cysteine residues. This exchange leads to a four-fold increase in catalytic activity.405 Other sulfhydryl reagents, such as l-fluoro-2,4-dinitrobenzene (FDNB), p-mercuri-benzoate (PMB), and 2-iodoacetamide, also activate liver D-fructose... 

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