Bolton—Hunter Reagent

Bolton-Hunter Reagent N-Succinimidyl-3-(4-hydroxyphenyl)propionate MW 263.26 

SF1PP is relatively insoluble in aqueous environments and must be dissolved in an organic solvent prior to addition to a reaction medium. Suggested solvents include dioxane and DMSO that are low in water content to avoid hydrolysis of the NHS ester. 

A water-soluble version of the original Bolton-Hunter reagent also is available, called sulfo-SHPP (Thermo Fisher). This compound contains a negatively charged sulfonate group on the NHS ring structure, which provides enough hydrophilicity to allow direct addition to aqueous reaction mediums. 

The following procedure describes the iodination process for the Bolton-Hunter reagent and its subsequent use for the radiolabeling of protein molecules. Modification of other macromolecules can be done using the same general method. For particular labeling applications, optimization of the level of iodine incorporation may have to be done to obtain the best specific radioactivity with retention of biological activity. 

SHPP is relatively insoluble in aqueous environments and must be dissolved in 

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Figure 12.7 The Bolton-Hunter reagent may be radioiodinated at its phenolic ring structure prior to reaction with an amine-containing molecule to form an amide bond modification.

Add to the iodination reaction 5 pi of DMF and 100 pi of benzene. Mix to extract the iodinated Bolton-Hunter reagent into the organic phase. 

Add the protein solution to the dried, iodinated Bolton-Hunter reagent. 

Remove excess labeled Bolton-Hunter reagent by gel filtration or dialysis. 

Figure 20.17 Bolton-Hunter reagent may be used to add radioactive iodine labels to antibody molecules by modification of amines.

Davies, C.E., and Palek, J. (1981) 1251-labeling of platelet proteins with Bolton-Hunter reagent. Anal. 

Salmain, M., Vessieres, A., Butler, I., and Jaouen, G. (1991) N-Succinimidyl (4-pentynoate)hexacarbon yldicobalt A transition-metal carbonyl complex having similar uses to the Bolton-Hunter reagent. Bioconjugate Chem. 2, 13-15. 

Bollworm/budworm complex, 5 9 Bolometers, 19 143-144 Bolton-Hunter reagent, 21 274 Boltzmann distribution, 14 657 26 1035 Boltzmann s constant, 26 1035 numerical value of, 24 434 Boltzmann s law, 14 662 Bolzano magnesium manufacturing process, 15 342... 

NHj 1 -Flnoro-2,4-dinitrobenzene Acetanhydride Bolton-Hunter reagent Dansylchloride Formaldehyde Maleic acid anhydride N-Snccinimidyl propionate Phenylisocyanate Sodium borohydride Succinic anhydride... 

Conjugation of a protein with the radiolabeled Bolton-Hunter reagent in the first place modifies lysine side chains. 

Bolton-Hunter reagent (N-succinimidyl 3(4-hydroxy-5- [ iodophenyl)propionate) (70-150TBq/mMol) in benzene 0.1 M borate buffer, pH 8.5 0.2 M glycine in Soln. B... 

Several applications of photoreactive peptides require the presence of a radionuclide to allow specific and sensitive detection of the photo-cross-linked conjugates. In several cases, radioiodination of tyrosyl moieties and radiolabeled Bolton-Hunter reagents have been used. However, the presence of a radiolabel within the benzophenone photophore is desirable, particularly when the objective is to identify the site of photo-insertion of benzophenone. To this end some radiolabeled, benzophenone-based compounds have been developed and used in peptide synthesis, in particular tritiated Phe(4-Bz) (Scheme 24)J2161 [1-14C-carboxy]-4-benzoylbenzoic acid,1221 and 4-benzoyl-(2,3-3H2)-dihydrocinnamic acid.[154l In addition, 4-(4-hydroxybenzoyl)phenylalanine (Scheme 25) has been directly radioiodinated with Na125I and Chloramine-T)151 ... 

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