Membrane proteins are crucial targets for cancer biomarker discovery and drug development. However, in addition to the inherent challenges of hydrophobicity and low abundance, complete membrane proteome coverage of clinical specimen is usually hindered by the requirement of large amount of starting materials. Toward comprehensive membrane proteomic profiling for small amounts of samples (10 μg), we developed high-pH reverse phase (Hp-RP) combined with stop-and-go extraction tip (StageTip) technique, as a fast (∼15 min.), sensitive, reproducible, high-resolution and multiplexed fractionation method suitable for accurate quantification of the membrane proteome. This approach provided almost 2-fold enhanced detection of peptides encompassing transmembrane helix (TMH) domain, as compared with strong anion exchange (SAX) and strong cation exchange (SCX) StageTip techniques. Almost 5000 proteins (∼60% membrane proteins) can be identified in only 10 μg of membrane protein digests, showing the superior sensitivity of the Hp-RP StageTip approach. The method allowed up to 9- and 6-fold increase in the identification of unique hydrophobic and hydrophilic peptides, respectively. The Hp-RP StageTip method enabled in-depth membrane proteome profiling of 11 lung cancer cell lines harboring different EGFR mutation status, which resulted in the identification of 3983 annotated membrane proteins. This provides the largest collection of reference peptide spectral data for lung cancer membrane subproteome. Finally, relative quantification of membrane proteins between Gefitinib-resistant and -sensitive lung cancer cell lines revealed several up-regulated membrane proteins with key roles in lung cancer progression.
Numerous biomolecules possess α-D-glucosamine as structural component. However, chemical glycosylations aimed at this backbone are usually not easily attained without generating the unwanted β-isomer. We report herein a versatile approach in affording full α-stereoselectivity built upon a carefully selected set of orthogonal protecting groups on a D-glucosaminyl donor. The excellent stereoselectivity provided by the protecting group combination was found independent of leaving groups and activators. With the trichloroacetimidate as the optimum donor leaving group, core skeletons of glycosylphosphatidyl inositol anchors, heparosan, heparan sulfate, and heparin were efficiently assembled. The orthogonal protecting groups were successfully manipulated to further carry out the total syntheses of heparosan tri- and pentasaccharides and heparin di-, tetra-, hexa-, and octasaccharide analogues. Using the heparin analogues, heparin-binding hemagglutinin, a virulence factor of Mycobacterium tuberculosis, was found to bind at least six sugar units with the interaction notably being entropically driven.
Fondaparinux, a synthetic pentasaccharide based on the heparin antithrombin-binding domain, is an approved clinical anticoagulant. Although it is a better and safer alternative to pharmaceutical heparins in many cases, its high cost, which results from the difficult and tedious synthesis, is a deterrent for its widespread use. The chemical synthesis of fondaparinux was achieved in an efficient and concise manner from commercially available D-glucosamine, diacetone α-D-glucose, and penta-O-acetyl-D-glucose. The method involves suitably functionalized building blocks that are readily accessible and employs shared intermediates and a series of one-pot reactions that considerably reduce the synthetic effort and improve the yield.
Despite significant efforts in the past decade towards complete mapping of the human proteome, 3564 proteins (neXtProt, 09-2014) are still “missing proteins”. Over one-third of these missing proteins are annotated as membrane proteins, owing to their relatively challenging accessibility with standard shotgun proteomics. Using non-small cell lung cancer (NSCLC) as a model study, we aim to mine missing proteins from disease-associated membrane proteome, which may be still largely under-represented. To increase identification coverage, we employed Hp-RP StageTip pre-fractionation of membrane-enriched samples from 11 NSCLC cell lines. Analysis of membrane samples from 20 pairs of tumor and adjacent normal lung tissue were incorporated to include physiologically expressed membrane proteins. Using multiple search engines (X!Tandem, Comet and Mascot) and stringent evaluation of FDR (MAYU and PeptideShaker), we identified 7702 proteins (66% membrane proteins) and 178 missing proteins (74 membrane proteins) with PSM-, peptide-, and protein-level FDR of 1%. Through multiple reaction monitoring (MRM) using synthetic peptides, we provided additional evidences for 8 missing proteins including 7 with transmembrane helix domains (TMH). This study demonstrates that mining missing proteins focused on cancer membrane sub-proteome can greatly contribute to map the whole human proteome. All data were deposited into ProteomeXchange with the identifier PXD002224.
Aromatic
polyimide (PI) derivatives have recently been investigated
as redox-active electrode materials for Li-ion batteries because of
their high thermal stability and thermo-oxidative stability complemented
by excellent solvent resistance, good electrical and mechanical properties,
and chemical resistance. In this work, we report two PI derivatives
from a newly synthesized 4,4′-diamino-3″,4″-dicyanotriphenylamine
(DiCN-TPA) monomer and two dianhydrides, pyromellitic dianhydride
(PMDA) and 1,4,5,8-naphthalenetetracarboxylic dianhydride (NTCDA);
designated as TPA-PMPI and TPA-NTCPI, respectively, as electrode materials
for Li-ion batteries. Characterizations of the PIs reveal excellent
thermal stability and bipolar property. The incorporation of DiCN-TPA
into the polymer structure resulted to a disordered chain arrangement,
thus giving high glass transition temperatures (T
g). Electrochemical performance tests reveal that TPA-NTCPI
cathode delivered a reversible specific capacity of 150 mAh g–1 at 0.1 A g–1 and exhibited a stability
up to 1000 cycles. On the other hand, TPA-PMPI anode delivered a high
specific capacity of up to 1600 mAh g–1 at 0.1 A
g–1 after 100 cycles. The electrochemical performance
of TPA-NTCPI cathode and TPA-PMPI anode are both among the best compared
with other reported aromatic PI-based electrodes. The long cycle lifetime
and excellent battery performance further suggest that TPA-NTCPI and
TPA-PMPI are promising organic electrode materials for next generation
Li-ion batteries.
The alkaline stability of different tethered amine functional groups of fuel cell anion exchange membranes (AEMs), namely, trimethyl amine (TMA), 1‐azabicyclo[2.2.2]octane (ABCO), 1,4‐diazabicyclo[2.2.2]octane (DABCO), and N‐methylpiperidine (NMP), is investigated using density functional theory (DFT). Among the amine functional groups investigated, ABCO emerged as the most stable exhibiting the highest energy of barrier (EOB) of 33.5 kcal/mol, while DABCO has the lowest EOB of 30.0 kcal/mol due to the presence of an additional electron‐withdrawing nitrogen. The calculated lowest unoccupied molecular orbital (LUMO) energy revealed the trend of increasing alkaline stability against nucleophilic attack, consistent with their measured barrier energies: DABCO < TMA < NMP < ABCO. Most importantly, the DFT calculations confirmed the proposed multistep AEM degradation mechanism via the detachment of the whole vinylbenzyl quaternary ammonium group through the following steps: (1) nucleophilic attack leading to the loss of aromaticity with subsequent transformation to a quinodimethane moiety, (2) detachment of the quinodimethane‐like intermediate from the polymer backbone by the attack of superoxide and/or peroxy radicals via oxidative cleavage, and (3) the rearomatisation of the reaction intermediate.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.