Site-selective chemical
conjugation of synthetic molecules to proteins
expands their functional and therapeutic capacity. Current protein
modification methods, based on synthetic and biochemical technologies,
can achieve site selectivity, but these techniques often require extensive
sequence engineering or are restricted to the N-
or C-terminus. Here we show the computer-assisted
design of sulfonyl acrylate reagents for the modification of a single lysine residue on native protein sequences. This
feature of the designed sulfonyl acrylates, together with the innate
and subtle reactivity differences conferred by the unique local microenvironment
surrounding each lysine, contribute to the observed regioselectivity
of the reaction. Moreover, this site selectivity was predicted computationally,
where the lysine with the lowest pKa was
the kinetically favored residue at slightly basic pH. Chemoselectivity
was also observed as the reagent reacted preferentially at lysine,
even in those cases when other nucleophilic residues such as cysteine
were present. The reaction is fast and proceeds using a single molar
equivalent of the sulfonyl acrylate reagent under biocompatible conditions
(37 °C, pH 8.0). This technology was demonstrated by the quantitative
and irreversible modification of five different proteins including
the clinically used therapeutic antibody Trastuzumab without prior
sequence engineering. Importantly, their native secondary structure
and functionality is retained after the modification. This regioselective
lysine modification method allows for further bioconjugation through
aza-Michael addition to the acrylate electrophile that is generated
by spontaneous elimination of methanesulfinic acid upon lysine labeling.
We showed that a protein–antibody conjugate bearing a site-specifically
installed fluorophore at lysine could be used for selective imaging
of apoptotic cells and detection of Her2+ cells, respectively. This
simple, robust method does not require genetic engineering and may
be generally used for accessing diverse, well-defined protein conjugates
for basic biology and therapeutic studies.
Crude oil and hydrocarbon fuel spills are a perennial threat to aquatic environments. Inexpensive and sustainable sorbents are needed to mitigate the ecological harm of this pollution. To address this need, this study features a low‐density polysulfide polymer that is prepared by the direct reaction of sulfur and used cooking oils. Because both sulfur and cooking oils are hydrophobic, the polymer has an affinity for hydrocarbons such as crude oil and diesel fuel and can rapidly remove them from seawater. Through simple mechanical compression, the oil can be recovered and the polymer can be reused in oil spill remediation. The polysulfide is unique because it is prepared entirely from repurposed waste: sulfur is a by‐product of the petroleum industry and used cooking oil can be used as a comonomer. In this way, sulfur waste from the oil industry is used to make an effective sorbent for combatting pollution from that same sector.
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