Justin M. Chalker

Allyl Sulfides Are Privileged Substrates in Aqueous Cross-Metathesis: Application to Site-Selective Protein Modification

Allyl sulfides undergo efficient cross-metathesis in aqueous media with Hoveyda-Grubbs second generation catalyst 1. The high reactivity of allyl sulfides in cross-metathesis was exploited in the first examples of cross-metathesis on a protein surface. S-Allylcysteine was incorporated chemically into the protein, providing the requisite allyl sulfide handle. Preliminary efforts to genetically incorporate S-allylcysteine into proteins are also reported.

A Convenient Catalyst for Aqueous and Protein Suzuki−Miyaura Cross-Coupling

A phosphine-free palladium catalyst for aqueous Suzuki-Miyaura cross-coupling is presented. The catalyst is active enough to mediate hindered, ortho-substituted biaryl couplings but mild enough for use on peptides and proteins. The Suzuki-Miyaura couplings on protein substrates are the first to proceed in useful conversions. Notably, hydrophobic aryl and vinyl groups can be transferred to the protein surface without the aid of organic solvent since the aryl- and vinylboronic acids used in the coupling are water-soluble as borate salts. The convenience and activity of this catalyst prompts use in both general synthesis and bioconjugation.

Olefin Cross-Metathesis on Proteins: Investigation of Allylic Chalcogen Effects and Guiding Principles in Metathesis Partner Selection

Olefin metathesis has recently emerged as a viable reaction for chemical protein modification. The scope and limitations of olefin metathesis in bioconjugation, however, remain unclear. Herein we report an assessment of various factors that contribute to productive cross-metathesis on protein substrates. Sterics, substrate scope, and linker selection are all considered. It was discovered during this investigation that allyl chalcogenides generally enhance the rate of alkene metathesis reactions. Allyl selenides were found to be exceptionally reactive olefin metathesis substrates, enabling a broad range of protein modifications not previously possible. The principles considered in this report are important not only for expanding the repertoire of bioconjugation but also for the application of olefin metathesis in general synthetic endeavors.

Sulfur-Limonene Polysulfide: A Material Synthesized Entirely from Industrial By-Products and Its Use in Removing Toxic Metals from Water and Soil.

Abstract A polysulfide material was synthesized by the direct reaction of sulfur and d‐limonene, by‐products of the petroleum and citrus industries, respectively. The resulting material was processed into functional coatings or molded into solid devices for the removal of palladium and mercury salts from water and soil. The binding of mercury(II) to the sulfur‐limonene polysulfide resulted in a color change. These properties motivate application in next‐generation environmental remediation and mercury sensing.

Rapid Vortex Fluidics: Continuous Flow Synthesis of Amides and Local Anesthetic Lidocaine

Thin film flow chemistry using a vortex fluidic device (VFD) is effective in the scalable acylation of amines under shear, with the yields of the amides dramatically enhanced relative to traditional batch techniques. The optimized monophasic flow conditions are effective in ≤80 seconds at room temperature, enabling access to structurally diverse amides, functionalized amino acids and substituted ureas on multigram scales. Amide synthesis under flow was also extended to a total synthesis of local anesthetic lidocaine, with sequential reactions carried out in two serially linked VFD units. The synthesis could also be executed in a single VFD, in which the tandem reactions involve reagent delivery at different positions along the rapidly rotating tube with in situ solvent replacement, as a molecular assembly line process. This further highlights the versatility of the VFD in organic synthesis, as does the finding of a remarkably efficient debenzylation of p-methoxybenzyl amines.

Green chemistry and polymers made from sulfur

Polymers are among the most important mass-produced materials on the planet, yet they are largely derived from a finite supply of petrochemicals. To ensure the sustainable production of polymers and functional materials, alternative feedstocks are required. This Perspective examines this challenge in the context of an emerging class of polymers made from elemental sulfur. Because sulfur is a by-product of the petroleum industry, converting it into useful polymers and related materials is an advance in waste valorisation. Additionally, co-polymerisation of sulfur with renewable monomers represents an additional contribution to sustainability. These reactions are often solvent free and benefit from full atom economy, futher augmenting their Green Chemistry credentials. Applications of these materials will be discussed, with a spotlight on environmental benefits. A forward looking assessment of the opportunities for using sulfur polymers in Green Chemistry is also included.

Laying Waste to Mercury: Inexpensive Sorbents Made from Sulfur and Recycled Cooking Oils

Abstract Mercury pollution threatens the environment and human health across the globe. This neurotoxic substance is encountered in artisanal gold mining, coal combustion, oil and gas refining, waste incineration, chloralkali plant operation, metallurgy, and areas of agriculture in which mercury‐rich fungicides are used. Thousands of tonnes of mercury are emitted annually through these activities. With the Minamata Convention on Mercury entering force this year, increasing regulation of mercury pollution is imminent. It is therefore critical to provide inexpensive and scalable mercury sorbents. The research herein addresses this need by introducing low‐cost mercury sorbents made solely from sulfur and unsaturated cooking oils. A porous version of the polymer was prepared by simply synthesising the polymer in the presence of a sodium chloride porogen. The resulting material is a rubber that captures liquid mercury metal, mercury vapour, inorganic mercury bound to organic matter, and highly toxic alkylmercury compounds. Mercury removal from air, water and soil was demonstrated. Because sulfur is a by‐product of petroleum refining and spent cooking oils from the food industry are suitable starting materials, these mercury‐capturing polymers can be synthesised entirely from waste and supplied on multi‐kilogram scales. This study is therefore an advance in waste valorisation and environmental chemistry.

Chemo- and Regioselective Lysine Modification on Native Proteins

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.

The mercury problem in artisanal and small-scale gold mining

Abstract Mercury‐dependent artisanal and small‐scale gold mining (ASGM) is the largest source of mercury pollution on Earth. In this practice, elemental mercury is used to extract gold from ore as an amalgam. The amalgam is typically isolated by hand and then heated—often with a torch or over a stove—to distill the mercury and isolate the gold. Mercury release from tailings and vaporized mercury exceed 1000 tonnes each year from ASGM. The health effects on the miners are dire, with inhaled mercury leading to neurological damage and other health issues. The communities near these mines are also affected due to mercury contamination of water and soil and subsequent accumulation in food staples, such as fish—a major source of dietary protein in many ASGM regions. The risks to children are also substantial, with mercury emissions from ASGM resulting in both physical and mental disabilities and compromised development. Between 10 and 19 million people use mercury to mine for gold in more than 70 countries, making mercury pollution from ASGM a global issue. With the Minamata Convention on Mercury entering force this year, there is political motivation to help overcome the problem of mercury in ASGM. In this effort, chemists can play a central role. Here, the problem of mercury in ASGM is reviewed with a discussion on how the chemistry community can contribute solutions. Introducing portable and low‐cost mercury sensors, inexpensive and scalable remediation technologies, novel methods to prevent mercury uptake in fish and food crops, and efficient and easy‐to‐use mercury‐free mining techniques are all ways in which the chemistry community can help. To meet these challenges, it is critical that new technologies or techniques are low‐cost and adaptable to the remote and under‐resourced areas in which ASGM is most common. The problem of mercury pollution in ASGM is inherently a chemistry problem. We therefore encourage the chemistry community to consider and address this issue that affects the health of millions of people.

Precise Probing of Residue Roles by Post-Translational β,γ-C,N Aza-Michael Mutagenesis in Enzyme Active Sites

Biomimicry valuably allows the understanding of the essential chemical components required to recapitulate biological function, yet direct strategies for evaluating the roles of amino acids in proteins can be limited by access to suitable, subtly-altered unnatural variants. Here we describe a strategy for dissecting the role of histidine residues in enzyme active sites using unprecedented, chemical, post-translational side-chain-β,γ C–N bond formation. Installation of dehydroalanine (as a “tag”) allowed the testing of nitrogen conjugate nucleophiles in “aza-Michael”-1,4-additions (to “modify”). This allowed the creation of a regioisomer of His (iso-His, Hisiso) linked instead through its pros-Nπ atom rather than naturally linked via C4, as well as an aza-altered variant aza-Hisiso. The site-selective generation of these unnatural amino acids was successfully applied to probe the contributing roles (e.g., size, H-bonding) of His residues toward activity in the model enzymes subtilisin protease from Bacillus lentus and Mycobacterium tuberculosis pantothenate synthetase.