Crosslinking reagents cleavable

Brandon, D.L. (1980) Studies of sheep red blood cell membranes, using cleavable crosslinking reagents. Cell. Mol. Biol. 26, 569-573. 

Control experiments showed that all the crosslinks were cleavable. No crosslinks were formed if the reagent was omitted, if photolysis was omitted, or if the reagent was first inactivated by treatment with ammonium acetate. These experiments proved that neither monofunctional cross-linking, nor disulfide interchange to give crosslinking in the dark occurred. If the membranes were solubilized in SDS before irradiation no cross-... 

AJ Lomant, G Fairbanks. Chemical probes of extended biological structures Synthesis and properties of the cleavable protein crosslinking reagent [35S] dithiobis (succinimidyl propionate). J Mol Biol 104 243, 1976. 

The differences within these families of reagents generally relate to the length of the spacer or bridging portion of the molecule. Occasionally, the bridging portion itself is designed to be cleavable by one of a number of methods (Chapter 8). The great majority of homobifunctional, sulfhydryl-reactive crosslinkers mentioned in the literature are not readily available from commercial sources and would have to be synthesized to make use of them. The ones listed in this section are obtainable from Thermo Fisher. 

Figure 6.2 The trifunctional reagent sulfo-SBED reacts with amine-containing bait proteins via its NHS ester side chain. Subsequent interaction with a protein sample and exposure to UV light can cause crosslink formation with a second interacting protein. The biotin portion provides purification or labeling capability using avidin or streptavidin reagents. The disulfide bond on the NHS ester arm provides cleavability using disulfide reductants, which effectively transfers the biotin label to an unknown interacting protein.

The reagents in current use for protein crosslinking have one chemically reactive and one photochemically reactive arm, often connected by a cleavable bridge for subsequent two-dimensional electrophoretic analyses. Chemical attachment is carried out first, and the crosslink is completed by photolysis. Many of these so-called heterobifunctional molecules are listed in Table 5.1 and some of them, conforming to the criteria outlined below,... 

In another case in which a number of exemplary control experiments were done, Markwell and Fox (1980) crosslinked the outer membranes of enveloped viruses with methyl 3-(p-azidophenyl)dithio]propionimidate. Virus (4 mg protein/ml) was reacted with the imidate (0.1 to 0.5 mM) at 0°C for 30 min at pH 8.5. The reaction was quenched with 50 mM ammonium acetate, 50 mM NEM (30 min, 25 °C), and the vims recovered by centrifugation. After irradiation the crosslinked polypeptides were examined in a two-dimensional SDS-polyacrylamide gel. One complication was that the crosslinking pattern had to be compared with a native pattern of disulfide linkages, and a reagent with a different cleavable crosslink may have been a better choice. As mentioned above, the analysis was simplified by the use of surface labeling. 

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