Protein S-acylation is a dynamic lipid post-translational modification that can modulate the localization and activity of target proteins. In humans, the installation of the lipid onto target proteins is catalyzed by a family of 23 Asp-His-His-Cys domain-containing protein acyltransferases (DHHC-PATs). DHHCs are increasingly recognized as critical players in cellular signaling events and in human disease. However, progress elucidating the functions and mechanisms of DHHC "writers" has been hampered by a lack of chemical tools to perturb their activity in live cells. Herein, we report the synthesis and characterization of cyano-myracrylamide (CMA), a broad-spectrum DHHC family inhibitor with similar potency to 2-bromopalmitate (2BP), the most commonly used DHHC inhibitor in the field. Possessing an acrylamide warhead instead of 2BP's α-halo fatty acid, CMA inhibits DHHC family proteins in cellulo while demonstrating decreased toxicity and avoiding inhibition of the S-acylation eraser enzymes -two of the major weaknesses of 2BP. Our studies show that CMA engages with DHHC family proteins in cells, inhibits protein S-acylation, and disrupts DHHC-regulated cellular events. CMA represents an improved chemical scaffold for untangling the complexities of DHHC-mediated cell signaling by protein S-acylation. File list (2)download file view on ChemRxiv SA8_V28 changes accepted.pdf (16.08 MiB) download file view on ChemRxiv SI_Revision4.pdf (37.14 MiB)

<div><div><div><p>Protein S-acylation is a dynamic lipid post-translational modification that can modulate the localization and activity of target proteins. In humans, the installation of the lipid onto target proteins is catalyzed by a family of 23 Asp-His-His-Cys domain-containing protein acyltransferases (DHHC-PATs). DHHCs are increasingly recognized as critical players in cellular signaling events and in human disease. However, progress elucidating the functions and mechanisms of DHHC “writers” has been hampered by a lack of chemical tools to perturb their activity in live cells. Herein, we report the synthesis and characterization of PATi, a pan- DHHC inhibitor more potent than 2-bromopalmitate (2BP), the most commonly used DHHC inhibitor in the field. Possessing an acrylamide warhead, PATi pairs its gain in potency with decreases in both toxicity and inhibition of the S-acylation eraser enzymes – two of the major weaknesses of 2BP. Our studies show that PATi engages with DHHC family proteins in cells, inhibits protein S-acylation, and disrupts DHHC-regulated cellular events. PATi represents an improved chemical tool for untangling the complexities of DHHC-mediated cell signaling by protein S-acylation.</p></div></div></div>

Bromination of high-pressure, high-temperature (HPHT) nanodiamond (ND) surfaces has not been explored and can open new avenues for increased chemical reactivity and diamond lattice covalent bond formation. The large bond dissociation energy of the diamond lattice–oxygen bond is a challenge that prevents new bonds from forming, and most researchers simply use oxygen-terminated NDs (alcohols and acids) as reactive species. In this work, we transformed a tertiary-alcohol-rich ND surface to an amine surface with ∼50% surface coverage and was limited by the initial rate of bromination. We observed that alkyl bromide moieties are highly labile on HPHT NDs and are metastable as previously found using density functional theory. The strong leaving group properties of the alkyl bromide intermediate were found to form diamond–nitrogen bonds at room temperature and without catalysts. This robust pathway to activate a chemically inert ND surface broadens the modalities for surface termination, and the unique surface properties of brominated and aminated NDs are impactful to researchers for chemically tuning diamond for quantum sensing or biolabeling applications.

Bromination of high-pressure high-temperature (HPHT) nanodiamond (ND) surfaces has not been explored and can open new avenues for increased chemical reactivity and diamond lattice covalent bond formation. The large bond dissociation energy of the diamond lattice-oxygen bond is a challenge that prevents new bonds from forming and most researchers simply use oxygen-terminated ND (alcohols and acids) as a reactive species. In this work, we transformed a tertiary alcohol-rich ND surface to an amine surface with 50% surface coverage and was limited by the initial rate of bromination. We observed that alkyl-bromide moieties are highly labile on NDs and are metastable as previously found using density functional theory. The instability of the bromine terminated ND is explained by steric hindrance and poor surface energy stabilization. The strong leaving group properties of the alkyl-bromide intermediate were found to form diamond-nitrogen bonds at room temperature and without catalysts. The chemical lability of the brominated ND surface led to efficient amination with NH3•THF at 298 K, and a catalyst-free Sonogashira-type reaction with an alkyne-amine produced an 11-fold increase in amination rate. Overlapping spectroscopies under inert, temperature-dependent and open-air conditions provided unambiguous chemical assignments. Amine-terminated NDs and folic acid were conjugated using sulfo-NHS/EDC coupling reagents to form amide bonds, confirming that standard amine chemistry remains viable. This work supports that a robust pathway exists to activate a chemically inert diamond surface at room temperature, which broadens the pathways of bond formation when a reactive alkyl-bromide surface is prepared. The unique surface properties of brominated and aminated nanodiamond reported here are impactful to researchers who wish to chemically tune diamond for quantum sensing applications or as an electron source for chemical transformations.

Surface chemistry of materials that host quantum bits such as diamond are an important avenue of exploration as quantum computation and quantum sensing platforms mature. Interfacing diamond in general, and nanoscale diamond (ND) in particular with silica is a potential route to integrate the quantum bit into a photonic device, fiber optic, cells or tissues with flexible functionalization chemistry. While silica growth on ND cores has been used successfully for quantum sensing and biolabeling, the surface mechanism to initiate growth was unknown. This report describes the surface chemistry responsible for silica bond formation on diamond and uses X-ray absorption spectroscopy (XAS) to probe the diamond surface chemistry and its electronic structure with increasing silica thickness. A modified Stöber (Cigler) method was used to synthesize 2–35 nm thick shells of SiO2 onto carboxylic acid rich ND cores and the diamond features and surface structure were characterized by overlapping techniques including electron microscopy. Importantly, we discovered that SiO2 growth on carboxylated NDs eliminates the presence of carboxylic acids and that basic ethanolic solutions converts the ND surface to an alcohol-rich surface prior to silica growth. The data supports a mechanism that alcohols on the ND surface generate silyl-ether (ND-O-Si-(OH)3) bonds due to rehydroxylation by ammonium hydroxide in ethanol. Additionally, resonant inelastic X-ray scattering (RIXS) maps produced by the transition edge sensor supports the chemical analysis provided by XAS. The suppression of the diamond electronic structure as a function of SiO2 thickness was observed, and the Auger electron escape depth was modeled using the NIST database for the Simulation of Electron Spectra for Surface Analysis (SESSA) to support our experimental results. Researchers using high-pressure high temperature (HPHT) NDs or any alcohol-terminated material (metal oxides, oxidized silicon carbide or cubic-boron nitride) for quantum sensing applications may exploit these results to design new core-shell quantum sensors with base-catalyzed reactions and metal oxide precursors.