Abstract Molecular imaging holds considerable promise for elucidating biological processes in normal physiology as well disease states, but requires noninvasive methods identifying analytes at sub‐micromolar concentrations. Particularly useful are genetically encoded, single‐protein reporters that harness the power of molecular biology to visualize specific processes, such have been conspicuously lacking vivo magnetic resonance (MRI). Herein, we report TEM‐1 β‐lactamase (bla) a reporter...

The success of mRNA-based therapeutics and vaccines is attributed to their rapid development, adaptability, scalable production. Modified ribonucleotides like N1-methylpseudouridine enhance stability reduce immunogenicity but were recently found induce cellular immunity off-target, +1 ribosomal frameshifted protein. We developed a new platform using cell-free translation (CFT) liquid chromatography-tandem mass spectrometry (MS) detect, characterize, quantify antigen proteins from mRNA...

Protein cage self-assembly enables encapsulation and sequestration of small molecules, macromolecules, nanomaterials for many applications in bionanotechnology. Notably, wild-type thermophilic ferritin from Archaeoglobus fulgidus (AfFtn) exists as a stable dimer four-helix bundle proteins at low ionic strength, the protein forms hollow assembly 24 protomers high strength (∼800 mM NaCl). This process can also be initiated by highly charged gold nanoparticles (AuNPs) solution, leading to...

Abstract A supramolecular strategy for detecting specific proteins in complex media by using hyperpolarized 129 Xe NMR is reported. cucurbit[6]uril (CB[6])‐based molecular relay was programmed three sequential equilibrium conditions designing a two‐faced guest (TFG) that initially binds CB[6] and blocks the CB[6]–Xe interaction. The protein analyte recruits TFG frees binding. TFGs containing CB[6]‐ carbonic anhydrase II (CAII)‐binding domains were synthesized one or two steps. X‐ray...

Genetically encoded magnetic resonance imaging (MRI) contrast agents enable non-invasive detection of specific biomarkers<italic>in vivo</italic>.

Abstract The success of mRNA-based therapeutics and vaccines can be attributed to their rapid development, adaptability new disease variants, scalable production. Modified ribonucleotides are often used in or enhance stability reduce immunogenicity. However, substituting uridine with N 1 -methylpseudouridine has recently been shown result +1 ribosomal frameshifting that induces cellular immunity the translated off-target protein. To accelerate vaccine it is critical have analytical methods...

Abstract Genetically encoded (GE) contrast agents detectable by magnetic resonance imaging (MRI) enable non‐invasive visualization of gene expression and cell proliferation at virtually unlimited penetration depths. Using hyperpolarized 129 Xe in combination with chemical exchange saturation transfer, an MR approach known as hyper‐CEST, enables ultrasensitive protein detection biomolecular imaging. GE MRI developed to date include nanoscale proteinaceous gas vesicles well the monomeric...

The regiospecific prenylation of an aromatic amino acid catalyzed by a dimethylallyl-l-tryptophan synthase (DMATS) is key step in the biosynthesis many fungal and bacterial natural products. DMATS enzymes share common "ABBA" fold with divergent active site contours that direct alternative C–C, C–N, C–O bond-forming trajectories. DMATS1 from Fusarium fujikuroi catalyzes reverse N-prenylation l-Trp generating allylic carbocation dimethylallyl diphosphate (DMAPP) then alkylates indole nitrogen...

Indole prenyltransferases catalyze the prenylation of l-tryptophan (l-Trp) and other indoles to produce a diverse set natural products in bacteria, fungi, plants, many which possess useful biological properties. Among this family enzymes, CymD from Salinispora arenicola catalyzes reverse N1 l-Trp, an unusual reaction given poor nucleophilicity indole nitrogen. utilizes dimethylallyl diphosphate (DMAPP) as prenyl donor, catalyzing dissociation leaving group followed by nucleophilic attack...

Cryptophane-based biosensors are promising agents for the ultrasensitive detection of biomedically relevant targets via 129Xe NMR. Dynamic light scattering revealed that cryptophanes form water-soluble aggregates tens to hundreds nanometers in size. Acridine orange fluorescence quenching assays allowed quantitation aggregation state, with critical concentrations ranging from 200 nM 600 nM, depending on cryptophane species solution. The addition excess carbonic anhydrase (CA) protein target a...

Dysregulation of cellular ribose uptake can be indicative metabolic abnormalities or tumorigenesis. However, analytical methods are currently limited for quantifying concentration in complex biological samples. Here, we utilize the highly specific recognition by ribose-binding protein (RBP) to develop a single-protein sensor detectable via sensitive NMR technique known as hyperpolarized 129Xe chemical exchange saturation transfer (hyper-CEST). We demonstrate that RBP, with tunable site and...

Abstract A supramolecular strategy for detecting specific proteins in complex media by using hyperpolarized 129 Xe NMR is reported. cucurbit[6]uril (CB[6])‐based molecular relay was programmed three sequential equilibrium conditions designing a two‐faced guest (TFG) that initially binds CB[6] and blocks the CB[6]–Xe interaction. The protein analyte recruits TFG frees binding. TFGs containing CB[6]‐ carbonic anhydrase II (CAII)‐binding domains were synthesized one or two steps. X‐ray...

Detection of protein–protein interactions (PPIs) is limited by current bioanalytical methods. Reconstitution TEM-1—promoted here cFos/cJun leucine zipper interaction—gives rise to sensitive ¹²⁹Xe NMR signal in bacterial cells.

Polyamines are small organic cations that essential for cellular function in all kingdoms of life. Polyamine metabolism is regulated by enzyme-catalyzed acetylation-deacetylation cycles a fashion similar to the epigenetic regulation histone eukaryotes. Bacterial polyamine deacetylases particularly intriguing, because these enzymes share fold and eukaryotic deacetylases. Recently, acetylpolyamine amidohydrolase from deep earth halophile

Abstract Molecular imaging holds considerable promise for elucidating biological processes in normal physiology as well disease states, but requires noninvasive methods identifying analytes at sub‐micromolar concentrations. Particularly useful are genetically encoded, single‐protein reporters that harness the power of molecular biology to visualize specific processes, such have been conspicuously lacking vivo magnetic resonance (MRI). Herein, we report TEM‐1 β‐lactamase (bla) a reporter...

Abstract P‐X‐α‐cyano‐β‐styrenyl esters (X = H, Cl, CF 3 , NO 2 ) of N‐benzyloxycarbonyl valine were prepared. The rate constant for amide formation with methyl valinate was assayed; it increases increasing electron attracting power the para substituent. N‐hydroxysuccinimidoyl ester and p‐H‐α‐cyano‐β‐styrenyl have comparable reactivities.

Die Danksagung in dieser Zuschrift muss wie folgt lauten: “This work was supported by NIH R01-GM097478 and CDMRP-LCRP Concept Award no. LC130824. The ITC purchased via S10-DO016260. We thank Drs. George Furst Jun Gu for assistance with NMR spectroscopy. Dr. David Christianson the CAII plasmid crystallography. This is based upon research conducted at Northeastern Collaborative Access Team beamlines, which are funded National Institute of General Medical Sciences from Institutes Health (P41...