Jingjing Wan

A Phospho-Directed Macroporous Alumina-Silica Nanoreactor with Multi-Functions

A phospho-directed nanoreactor with multiple functions is reported. Alumina-functionalized macroporous ordered silica foams (Al-MOSF) have been developed with large pore size, high pore volume (1.6 cm(3)/g), and a surface area of 186 m(2)/g rich in coordination unsaturated Al species, which can be used as phospho-directed nanoreactors for integrated in situ digestion and in situ phosphoisolation. By directly adding Al-MOSF to the conventional in-solution digestion system, both enzymes and proteins are quickly enriched in the macropores of the reactor to achieve a fast proteolysis without increasing the enzyme/protein concentration or using a preimmobilization process, thus the digestion time and the cost can be greatly reduced. Meanwhile, due to the chemo-affinity between alumina and phosphor groups, the Al-MOSF reactor can in situ isolate specific products of the enzymatic reaction (i.e., phosphopeptides) and release the nonspecific peptides to the solution. This strategy is simple, efficient, and successfully applied in the detection of phosphoproteins in real samples.

Selenoether oxytocin analogues have analgesic properties in a mouse model of chronic abdominal pain

Poor oral availability and susceptibility to reduction and protease degradation is a major hurdle in peptide drug development. However, drugable receptors in the gut present an attractive niche for peptide therapeutics. Here we demonstrate, in a mouse model of chronic abdominal pain, that oxytocin receptors are significantly upregulated in nociceptors innervating the colon. Correspondingly, we develop chemical strategies to engineer non-reducible and therefore more stable oxytocin analogues. Chemoselective selenide macrocyclization yields stabilized analogues equipotent to native oxytocin. Ultra-high-field nuclear magnetic resonance structural analysis of native oxytocin and the seleno-oxytocin derivatives reveals that oxytocin has a pre-organized structure in solution, in marked contrast to earlier X-ray crystallography studies. Finally, we show that these seleno-oxytocin analogues potently inhibit colonic nociceptors both in vitro and in vivo in mice with chronic visceral hypersensitivity. Our findings have potentially important implications for clinical use of oxytocin analogues and disulphide-rich peptides in general.

TiO2‐Modified Macroporous Silica Foams for Advanced Enrichment of Multi‐Phosphorylated Peptides

Enriching peptides: Novel TiO(2)-modified macroporous materials (Ti-MOSF, see figure) have been synthesized with high surface area, large pore volume, and functional surfaces that are rich in coordinatively unsaturated Ti(IV) species, which can be applied in the specific extraction of phosphopeptides and which show a preferential capture of multi-phosphorylated peptides with low detection limits and high selectivity.

Macroporous Materials as Novel Catalysts for Efficient and Controllable Proteolysis

A novel nanopore based digestion strategy has been developed by directly adding a macroporous material as catalyst to the conventional in-solution reaction system. Without increasing the enzyme or protein concentrations, this simple digestion approach exhibits high proteolysis efficiency and selectivity due to the in situ fast adsorption of both enzymes and proteins from bulk solution into the macropores of the catalysts, where the target substrates and enzymes are greatly concentrated and confined in the nanospace to realize a quick digestion. Based on the electrostatic interaction matching between the biomolecules and catalysts, selective extraction and digestion of proteins with different isoelectric points can be achieved by adjusting the surface charge of the catalysts. This nanoporous reaction system has been successfully applied to the analysis of a complex biological sample, where 293 proteins are identified, while only 100 proteins are obtained by the standard overnight in-solution digestion. The present nanospace confined digestion strategy will lead to promising advances not only in proteomics but also in other applications where enzymatic reactions are involved.

A Smart Glycol‐Directed Nanodevice from Rationally Designed Macroporous Materials

We have developed a smart nanodevice for the highly efficient and selective detection of glycoproteins. This polyfunctional device is fabricated through the rational functionalization of macroporous silica foam (MOSF) materials with a boron species (B-MOSF) and amino groups (NH(2)-MOSF), and then the integration of MOSF, B-MOSF and NH(2)-MOSF materials. In such a device, a macroporous structure with very large-pore sizes (diameters approximately 100 nm) and high-pore volumes (>0.65 cm(3) g(-1)) is advantageous to efficiently fasten the enzymatic reaction. The targeted specific glycopeptides of the products can be selectively isolated and enriched in B-MOSF through the chemo-affinity between boronic acid and glycol groups, while the non-specific peptides are released to the solutions, or further purified by MOSF and NH(2)-MOSF, which have opposite charges. As a result, the protein digestion and glycol-peptide isolation can be simultaneously achieved in the functionalized macroporous materials in one step, which is a great advantage compared to conventional multi-procedure and time-consuming techniques.

Functionalized Periodic Mesoporous Organosilicas for Enhanced and Selective Peptide Enrichment

The analysis of peptides by the mass spectrometry (MS) technique is important in modern life science. The enrichment of peptides can increase the detection efficiency and is sometimes indispensable for collecting the information on proteins with low-abundance. Herein, we first report that functionalized periodic mesoporous organosilica (PMO) materials have a superior peptide enrichment property. It is demonstrated that the PMO materials with an organo-bridged (-CH(2)-) hybrid wall composition display a highly enhanced peptide enrichment ability compared to the pure silica material (SBA-15) with similar mesostructured parameters and morphology. More importantly, by surface modification of PMO with amino groups (denoted NH(2)-PMO), PMO and NH(2)-PMO with opposite charged surfaces (-25.2 and +39.0 mV, respectively) show selective affinities for positively and negatively charged peptides, respectively. By directly adding PMO, NH(2)-PMO as well as pure silica materials to the peptides solution with a low concentration (1-2 fmol/microL), 36 and 28 peptides can be detected from the BSA digestion in the presence of PMO and NH(2)-PMO, respectively, while only 6 and 4 are monitored in the case of SBA-15 enrichment and from solution without enrichment, respectively. Moreover, 69.4% (25 of 36) of enriched peptides by PMO have pI > or = 6 and 80% (21 of 28) of enriched peptides by NH(2)-PMO possess pI < or = 6. Combining the results from the NH(2)-PMO and PMO enrichment together, 51 peptides can be identified with a MOWSE score of 333. It is also noted that similar conclusions can also be obtained from the peptides solution originated from other proteins. This might be an important contribution to the understanding of the interaction between peptides and porous hosts, and the proposed method is promising for the development of both material science and biotechnology.

MALDI In-Source Photooxidation Reactions for Online Peptide Tagging†

F o r MALDI-MS analysis , thephotoelectrode is coated with the sample and a matrixoverlayer (a -cyano-4-hydroxycinnamic acid ; CHCA) isadded to assist light-energy absorption and protonation ofthe sample . One advantage of the photoelectrode-modifiedplate is that it enables the determination of the oxidation orreduction products of a given molecule . If this oxidation/reduction induces fragmentation of the analytes , the photo-electrode generates in-source decay upon irradiation. Alter -natively ,the oxidation/reduction products may in turn furtherreact with other molecules , and all the products of theseelectron-transfer chain reactions can be directly studied bymass spectrometry .T o illustrate this principle ,w e use the addition reaction ofoxidized hydroquinone on cysteine-containing peptides ,which has previously been demonstrated with electrosprayionization.

α-Conotoxin Dendrimers Have Enhanced Potency and Selectivity for Homomeric Nicotinic Acetylcholine Receptors

Covalently attached peptide dendrimers can enhance binding affinity and functional activity. Homogenous di- and tetravalent dendrimers incorporating the α7-nicotinic receptor blocker α-conotoxin ImI (α-ImI) with polyethylene glycol spacers were designed and synthesized via a copper-catalyzed azide-alkyne cycloaddition of azide-modified α-ImI to an alkyne-modified polylysine dendron. NMR and CD structural analysis confirmed that each α-ImI moiety in the dendrimers had the same 3D structure as native α-ImI. The binding of the α-ImI dendrimers to binding protein Ac-AChBP was measured by surface plasmon resonance and revealed enhanced affinity. Quantitative electrophysiology showed that α-ImI dendrimers had ∼100-fold enhanced potency at hα7 nAChRs (IC50 = 4 nM) compared to native α-ImI (IC50 = 440 nM). In contrast, no significant potency enhancement was observed at heteromeric hα3β2 and hα9α10 nAChRs. These findings indicate that multimeric ligands can significantly enhance conotoxin potency and selectivity at homomeric nicotinic ion channels.

Plasmonic silver nanoshells for drug and metabolite detection

In-vitro metabolite and drug detection rely on designed materials-based analytical platforms, which are universally used in biomedical research and clinical practice. However, metabolic analysis in bio-samples needs tedious sample preparation, due to the sample complexity and low molecular abundance. A further challenge is to construct diagnostic tools. Herein, we developed a platform using silver nanoshells. We synthesized SiO2@Ag with tunable shell structures by multi-cycled silver mirror reactions. Optimized nanoshells achieved direct laser desorption/ionization mass spectrometry in 0.5 μL of bio-fluids. We applied these nanoshells for disease diagnosis and therapeutic evaluation. We identified patients with postoperative brain infection through daily monitoring and glucose quantitation in cerebrospinal fluid. We measured drug distribution in blood and cerebrospinal fluid systems and validated the function of blood-brain/cerebrospinal fluid-barriers for pharmacokinetics. Our work sheds light on the design of materials for advanced metabolic analysis and precision diagnostics.Preparation of samples for diagnosis can affect the detection of biomarkers and metabolites. Here, the authors use a silver nanoparticle plasmonics approach for the detection of biomarkers in patients as well as investigate the distribution of drugs in serum and cerebral spinal fluid.

Peptide-Decorated Dendrimers and Their Bioapplications

Peptide-decorated dendrimers (PDDs) are a class of spherical, regular, branched polymers that are modified by peptides covalently attached to their surface. PDDs have been used as protein mimetics, novel biomaterials, and in a wide range of biomedical applications. Since their design and development in the late eighties, poly-l-lysine has been a preferred core structure for PDDs. However, numerous recent innovations in polymer synthesis and ligation chemistry have re-energized the field and led to the emergence of well-defined peptide dendrimers with more diverse core structures and functions. This Minireview highlights the development of PDDs driven by significantly improved ligation chemistry incorporating structurally well-defined peptides and the emerging use of PDDs in imaging and drug development.