ChemBioChem
10.1002/cbic.201900692
COMMUNICATION
1
3
a
Keywords: protein modification • N-terminal modification •
Yield of TA4C
MOPS (10 mol%)
1
: 93%
10: 81%
triazolecarbaldehyde • Dimroth rearrangement
DMSO
1
4: quant. 15: 95%
9
9 ℃, 30 min
[
[
[
[
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MOPS buffer
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37 ℃, 16 h
Protein
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[
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Figure 5. N-terminal modification of RNase using TA4C reagents prepared in a
one-step process via a Dimroth rearrangement. Deconvoluted ESI-MS spectra
of modified proteins are shown. Peaks for unmodified RNase are marked with
open circles and species corresponding to the correctly modified protein are
labeled with filled circles. Conditions for Dimroth rearrangement: amine
precursor (100 mM), 13 (100 mM), MOPS (10 mol%), DMSO at 99 °C for 30
min. Conditions for protein modification: protein (50 M), 1, 10, 14, 15 (ca. 10
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4015-4019.
1
b
Dimroth rearrangement was analyzed by H NMR measurements. The relative
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derivatives (TA4C), which are synthesized in an efficient CuAAC
reaction. A series of the TA4C reagents with functional moieties such as
fluorophores, biotin, and polyethylene glycol enables highly specific N-
terminal modification of various proteins. Furthermore, we
demonstrated direct preparation of the TA4C moiety from an amine
compound having a functional group through a Dimroth rearrangement
and used the product as a reagent for N-terminal protein modification.
Our results indicate that protein modification using a TA4C derivative
will serve as a simple and powerful method for conjugation of a wide
range of molecules and biomolecules as payloads at the N-terminus to
functionalize proteins in only two steps: (i) preparation of TA4C and (ii)
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for proteins is expected to accelerate the generation of designed proteins
loaded with multiple chemical or biomolecular entities at the N-terminus
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technologies using our TA4C reagent for protein conjugates such as
antibody-drug conjugates in medical and diagnostic applications are
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