Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Facile folding of insulin variants bearing a prosthetic C-peptide prepared by ?-ketoacid-hydroxylamine (KAHA) ligation.

ABSTRACT: The chemical synthesis of insulin is an enduring challenge due to the hydrophobic peptide chains and construction of the correct intermolecular disulfide pattern. We report a new approach to the chemical synthesis of insulin using a short, traceless, prosthetic C-peptide that facilitates the formation of the correct disulfide pattern during folding and its removal by basic treatment. The linear precursor is assembled by an ester forming ?-ketoacid-hydroxylamine (KAHA) ligation that provides access to the linear insulin precursors in good yield from two readily prepared segments. This convergent and flexible route provides access to various human, mouse, and guinea pig insulins containing a single homoserine mutation that shows no detrimental effect on the biological activities.

SUBMITTER: Boross GN

PROVIDER: S-EPMC6243641 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature

ACCESS DATA

Json Xml

Publications

Facile folding of insulin variants bearing a prosthetic C-peptide prepared by α-ketoacid-hydroxylamine (KAHA) ligation.

Boross Gábor N GN Shimura Satomi S Besenius Melissa M Tennagels Norbert N Rossen Kai K Wagner Michael M Bode Jeffrey W JW

Chemical science 20180911 44

The chemical synthesis of insulin is an enduring challenge due to the hydrophobic peptide chains and construction of the correct intermolecular disulfide pattern. We report a new approach to the chemical synthesis of insulin using a short, traceless, prosthetic C-peptide that facilitates the formation of the correct disulfide pattern during folding and its removal by basic treatment. The linear precursor is assembled by an ester forming α-ketoacid-hydroxylamine (KAHA) ligation that provides acce ...[more]

PMID: 30542587

Similar Datasets

Chemical synthesis of a homoserine-mutant of the antibacterial, head-to-tail cyclized protein AS-48 by ?-ketoacid-hydroxylamine (KAHA) ligation.

Project description:An antibacterial cyclic AS-48 protein was chemically synthesized by ?-ketoacid-hydroxylamine (KAHA) ligation. Initial challenges associated with the exceptionally hydrophobic segments arising from the amphiphilic nature of the protein were resolved by the development of bespoke reaction conditions for hydrophobic segments, using hexafluoroisopropanol (HFIP) as a co-solvent. The synthetic protein displays similar biological activity and properties to those of the native protein. To support the current understanding of its antibacterial mode of action, we demonstrate the ability of AS-48 to be incorporated into synthetic multilamellar vesicles (MLVs).

| S-EPMC5434751 | biostudies-literature

Facile synthesis of insulin fusion derivatives through sortase A ligation.

Project description:Insulin derivatives such as insulin detemir and insulin degludec are U.S. Food and Drug Administration (FDA)-approved long-acting insulin currently used by millions of people with diabetes. These derivatives are modified in C-terminal B29 lysine to retain insulin bioactivity. New and efficient methods for facile synthesis of insulin derivatives may lead to new discovery of therapeutic insulin. Herein, we report a new method using sortase A (SrtA)-mediated ligation for the synthesis of insulin derivatives with high efficiency and functional group tolerance in the C-terminal B chain. This new insulin molecule (Ins-SA) with an SrtA-recognizing motif can be conjugated to diverse groups with N-terminal oligoglycines to generate new insulin derivatives. We further demonstrated that a new insulin derivative synthesized by this SrtA-mediated ligation shows strong cellular and in vivo bioactivity. This enzymatic method can therefore be used for future insulin design and development.

| S-EPMC8463260 | biostudies-literature

Primitive templated catalysis of a peptide ligation by self-folding RNAs.

Project description:RNA-polypeptide complexes (RNPs), which play various roles in extant biological systems, have been suggested to have been important in the early stages of the molecular evolution of life. At a certain developmental stage of ancient RNPs, their RNA and polypeptide components have been proposed to evolve in a reciprocal manner to establish highly elaborate structures and functions. We have constructed a simple model system, from which a cooperative evolution system of RNA and polypeptide components could be developed. Based on the observation that several RNAs modestly accelerated the chemical ligation of the two basic peptides. We have designed an RNA molecule possessing two peptide binding sites that capture the two peptides. This designed RNA can also accelerate the peptide ligation. The resulting ligated peptide, which has two RNA-binding sites, can in turn function as a trans-acting factor that enhances the endonuclease activity catalyzed by the designed RNA.

| S-EPMC2677871 | biostudies-other

Fluorinated polymer surfactants bearing an alternating peptide skeleton prepared by three-component polycondensation.

Project description:A new species of fluorinated polymer surfactant was developed by three component polycondensation analogous to Ugi four-component condensation. The surfactant exhibited unique surface properties, which made cellulose-based materials hydrophobic and decreased the surface tension of CHCl3. It turned out that the polymer forms micelles in CHCl3.

| S-EPMC9078376 | biostudies-literature

Abiotic hydroxylamine nitrification involving manganese- and iron-bearing minerals.

Project description:Hydroxylamine (NH2OH) undergoes biotic and abiotic transformation processes in soil, producing nitrous oxide gas (N2O(g)). Little is known about the magnitude of the abiotic chemical processes in the global N cycle, and the role of abiotic nitrification is still neglected in most current nitrogen trace gas studies. The abiotic fate of NH2OH in soil systems is often focused on transition metals including manganese (Mn) and iron (Fe), and empirical correlations of nitrogen residual species including nitrite (NO2-), nitrate (NO3-), and N2O(g). In this study, abiotic NH2OH nitrification by well-characterized manganese (Mn)- and iron (Fe)-bearing minerals (pyrolusite, amorphous MnO2(s), goethite, amorphous FeOOH(s)) was investigated. A nitrogen mass balance analysis involving NH2OH, and the abiotic nitrification residuals, N2O(g), N2O(aq), NO2-, NO3-, was used, and specific reactions and mechanisms were investigated. Rapid and complete NH2OH nitrification occurred (4-5 h) in the presence of pyrolusite and amorphous MnO2(s), achieving a 95-96% mass balance of N byproducts. Conversely, NH2OH nitrification was considerably slower by amorphous FeOOH(s) (14.5%) and goethite (1.1%). Direct reactions between the Mn- and Fe-bearing mineral species and NO2- and NO3- were not detected. Brunauer-Emmett-Teller surface area and energy dispersive X-ray measurements for elemental composition were used to determine the specific concentrations of Mn and Fe. Despite similar specific concentrations of Mn and Fe in crystalline and amorphous minerals, the rate of NH2OH nitrification was much greater in the Mn-bearing minerals. Results underscore the intrinsically faster NH2OH nitrification by Mn minerals than Fe minerals.

| S-EPMC7286054 | biostudies-literature

Biomimetic synthesis of lispro insulin via a chemically synthesized "mini-proinsulin" prepared by oxime-forming ligation.

Project description:Here we report a proof-of-principle study demonstrating the efficient folding, with concomitant formation of the correct disulfides, of an isolated polypeptide insulin precursor of defined covalent structure. We used oxime-forming chemical ligation to introduce a temporary "chemical tether" to link the N-terminal residue of the insulin A chain to the C-terminal residue of the insulin B chain; the tether enabled us to fold/form disulfides with high efficiency. Enzymatic removal of the temporary chemical tether gave mature, fully active insulin. This chemical tethering principle could form the basis of a practical, high yield total synthesis of insulin and analogues.

| S-EPMC3690821 | biostudies-literature

Branched-chain ketoacid overload inhibits insulin action in the muscle.

Project description:Branched-chain α-keto acids (BCKAs) are catabolites of branched-chain amino acids (BCAAs). Intracellular BCKAs are cleared by branched-chain ketoacid dehydrogenase (BCKDH), which is sensitive to inhibitory phosphorylation by BCKD kinase (BCKDK). Accumulation of BCKAs is an indicator of defective BCAA catabolism and has been correlated with glucose intolerance and cardiac dysfunction. However, it is unclear whether BCKAs directly alter insulin signaling and function in the skeletal and cardiac muscle cell. Furthermore, the role of excess fatty acids (FAs) in perturbing BCAA catabolism and BCKA availability merits investigation. By using immunoblotting and ultra-performance liquid chromatography MS/MS to analyze the hearts of fasted mice, we observed decreased BCAA-catabolizing enzyme expression and increased circulating BCKAs, but not BCAAs. In mice subjected to diet-induced obesity (DIO), we observed similar increases in circulating BCKAs with concomitant changes in BCAA-catabolizing enzyme expression only in the skeletal muscle. Effects of DIO were recapitulated by simulating lipotoxicity in skeletal muscle cells treated with saturated FA, palmitate. Exposure of muscle cells to high concentrations of BCKAs resulted in inhibition of insulin-induced AKT phosphorylation, decreased glucose uptake, and mitochondrial oxygen consumption. Altering intracellular clearance of BCKAs by genetic modulation of BCKDK and BCKDHA expression showed similar effects on AKT phosphorylation. BCKAs increased protein translation and mTORC1 activation. Pretreating cells with mTORC1 inhibitor rapamycin restored BCKA's effect on insulin-induced AKT phosphorylation. This study provides evidence for FA-mediated regulation of BCAA-catabolizing enzymes and BCKA content and highlights the biological role of BCKAs in regulating muscle insulin signaling and function.

| S-EPMC7667962 | biostudies-literature

Trimodal chromatography enables facile peptide fractionation

Project description:We evaluated the utility of a trimodal stationary phase (Trinity P1) in offline peptide fractionation, and found it performed similarly to ion exchange or high pH reversed phase based techniques. It is orthogonal to low pH reversed phase and uses MS compatible mobile phase solvents.

2017-08-07 | PXD006188 | Pride

Facile Synthesis of Asymmetric aza-Boron Dipyrromethene Analogues Bearing Quinoxaline Moiety.

Project description:An asymmetric aza-BODIPY analogue bearing quinoxaline moiety was synthesized via a titanium tetrachloride-mediated Schiff-base-forming reaction of 6,7-dimethyl-1,4-dihydroquinoxaline-2,3-dione and benzo[d]thiazol-2-amine. This novel aza-BODIPY analogue forms a complementary hydrogen-bonded dimer due to the quinoxaline moiety in the crystal structure. It also shows intense absorption and fluorescence, with fluorescence quantum yields close to unity. The electrochemical measurements and the DFT calculations revealed the presence of the low-lying HOMO, which benefits their potential applications as an electron-transporting material.

| S-EPMC10745853 | biostudies-literature

Construction of diverse peptide structural architectures <i>via</i> chemoselective peptide ligation.

Project description:Herein, we report the development of a facile synthetic strategy for constructing diverse peptide structural architectures via chemoselective peptide ligation. The key advancement involved is to utilize the benzofuran moiety as the peptide salicylaldehyde ester surrogate, and Dap-Ser/Lys-Ser dipeptide as the hydroxyl amino functionality, which could be successfully introduced at the side chain of peptides enabling peptide ligation. With this method, the side chain-to-side chain cyclic peptide, branched/bridged peptides, tailed cyclic peptides and multi-cyclic peptides have been designed and successfully synthesized with native peptidic linkages at the ligation sites. This strategy has provided an alternative strategic opportunity for synthetic peptide development. It also serves as an inspiration for the structural design of PPI inhibitors with new modalities.

| S-EPMC8153220 | biostudies-literature

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data