Abstract
The advantages of using amino-directed photoprobes to couple native fragments, obtained by enzymatic digestion with trypsin, to protein carriers to prepare peptide-conjugates is described. The following photoprobe reagents were investigated:N-hydroxysuccinimidylp-azidobenzoate,N-hydroxysuccinimidyl ester ofp-azidobenzoylglycine,N-hydroxysuccinimidylp-benzoylbenzoate, and pentachlorophenyl ester ofp-benzoylbenzoyl glycine or the symmetric anhydride ofp-benzoylbenzoylglycine. These reagents modify only the NH2-terminal amino group and/or COOH-terminal ε-amino group of lysine of the tryptic fragments. Since the photoprobe is inert until photolysis, the probe-modified native fragment can be readily purified by high-performance liquid chromatography before cross-linking to the carrier molecule. The benzophenone photoprobes were shown to give the highest incorporation of peptide onto the protein carrier.
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Parker, J.M.R., Hodges, R.S. II. Photoaffinity probes provide a general method to prepare peptide-conjugates from native protein fragments. J Protein Chem 3, 479–489 (1984). https://doi.org/10.1007/BF01025066
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DOI: https://doi.org/10.1007/BF01025066