High-Performance Graphene-Titania Platform for Detection of Phosphopeptides in Cancer Cells

Tang, Lena Ai Ling; Wang, Junzhong; Lim, Teck Kwang; Bi, Xuezhi; Lee, Wong Cheng; Lin, Qingsong; Chang, Young‐Tae; Lim, Chwee Teck; Loh, Kian Ping

doi:10.1021/ac301119r

Cited by 64 publications

(40 citation statements)

References 52 publications

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“…The detailed methods used for LC-MS/MS were described previously (26). Briefly, separation of the iTRAQ-labeled peptides was carried out on an Eksigent nanoLC Ultra and ChiPLC-nanoflex (Eksigent, Dublin, CA) in the "Trap Elute" configuration.…”

Section: Protein Identification and Quantificationnano-lc Electrospramentioning

confidence: 99%

A Quantitative Chemical Proteomics Approach to Profile the Specific Cellular Targets of Andrographolide, a Promising Anticancer Agent That Suppresses Tumor Metastasis

Wang

Tan

Nguyễn

et al. 2014

Molecular & Cellular Proteomics

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Drug target identification is a critical step toward understanding the mechanism of action of a drug, which can help one improve the drug's current therapeutic regime and expand the drug's therapeutic potential. However, current in vitro affinity-chromatography-based and in vivo activity-based protein profiling approaches generally face difficulties in discriminating specific drug targets from nonspecific ones. Here we describe a novel approach combining isobaric tags for relative and absolute quantitation with clickable activity-based protein profiling to specifically and comprehensively identify the protein targets of andrographolide (Andro), a natural product with known anti-inflammation and anti-cancer effects, in live cancer cells. We identified a spectrum of specific targets of Andro, which furthered our understanding of the mechanism of action of the drug. Our findings, validated through cell migration and invasion assays, showed that Andro has a potential novel application as a tumor metastasis inhibitor. Moreover, we have unveiled the target binding mechanism of Andro with a combination of drug analog synthesis, protein engineering, and mass-spectrometry-based approaches and determined the drugbinding sites of two protein targets, NF-B and actin. Molecular & Cellular Proteomics 13: 10.1074/mcp. M113.029793, 876-886, 2014.As most drugs exert pharmacological effects by interacting with their target proteins, the identification of these target proteins is a critical step in unraveling the mechanisms of drug action. It is also imperative for our understanding of the pharmacodynamics of a known drug, suggesting potentially unrevealed actions and thus refining future clinical applications of the substance. Traditional approaches used to identify protein targets of a drug typically utilize immobilized drug affinity chromatography coupled with mass spectrometry (MS) 1 (1, 2). These methods can be applied to cell lysates, but not in an in vivo setting, because of the requirement of a solid support. In vitro target profiling might not accurately reflect the drug's actions in the in vivo physiological environment. To overcome this limitation, several groups have used activity-based protein profiling (ABPP) combined with bio-orthogonal click chemistry to identify drug targets both in vitro and in vivo (supplemental Fig. S1) (3-15). ABPP probes exert their functions via covalent reactions with the target proteins or photoaffinity-based labeling via the incorporation of photoreactive groups. With the increasing sensitivity of modern MS platforms, low-abundance protein targets can be successfully identified. Although both conventional affinity chromatography and recent ABPP-based methods allow us to detect a set of candidate protein targets for a drug, it remains difficult to 1 The abbreviations used are: MS, mass spectrometry; ABPP, activity-based protein profiling; ICABPP, clickable activity-based protein profiling; iTRAQ, isobaric tags for relative and absolute quantitation; DMSO, dimethyl sulfoxide; Andro, androgr...

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Section: Protein Identification and Quantificationnano-lc Electrospramentioning

confidence: 99%

A Quantitative Chemical Proteomics Approach to Profile the Specific Cellular Targets of Andrographolide, a Promising Anticancer Agent That Suppresses Tumor Metastasis

Wang

Tan

Nguyễn

et al. 2014

Molecular & Cellular Proteomics

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Section: Discussionmentioning

confidence: 99%

“…Loh's group developed a G-titania (G-TiO 2 ) platform for selective extraction of phosphopeptides from cancer cells [122]. These functionalized platforms were selectively conjugated with phosphopeptides, thereby increasing and expanding the library of phosphopeptides in cancer cells, which is useful in profiling disease-related signaling pathways.…”

Section: Nanomaterials-based Maldi-ms For Bacterial Identification Anmentioning

confidence: 99%

Nanomaterial-based miniaturized extraction and preconcentration techniques coupled to matrix-assisted laser desorption/ionization mass spectrometry for assaying biomolecules

Kailasa

2015

TrAC Trends in Analytical Chemistry

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“…Hybridization with other nanomaterials to obtain better selectivity for specific analytes (e.g., with TiO 2 for phosphopeptides [97][98][99][100] Yan et al [120] reported an IMAC (immobilized metal ion affinity chromatography) material with polydopamine coated on the surface of graphene and functionalized with titanium ions (Ti 4+ -G@PD) as an immobilized metal ion affinity chromatography platform for phosphoproteome followed by MALDI-TOF MS. In this approach, the application of the Ti …”

Section: Graphenementioning

confidence: 99%

Recent progress in application of carbon nanomaterials in laser desorption/ionization mass spectrometry

et al. 2016

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Carbon nanomaterials have attracted great interest over past decades owing to their unique physical properties, versatile functionalization chemistry, and biological compatibility. In this article, we review recent progress in application of carbon nanomaterials in laser desorption/ionization mass spectrometry (LDI MS). Various types of carbon nanomaterials, including fullerenes, carbon nanotubes, graphene, carbon nanodots, nanodiamond, nanofibers, nanohorns, and their derivative forms, are involved. The applications of these materials as new matrices or probes in matrix-assisted or surface-enhanced laser desorption/ ionization mass spectrometry (MALDI or SELDI MS) are discussed. Finally, we summarize current challenges and give our perspectives on the future of applications of carbon nanomaterials in LDI MS.

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High-Performance Graphene-Titania Platform for Detection of Phosphopeptides in Cancer Cells

Cited by 64 publications

References 52 publications

A Quantitative Chemical Proteomics Approach to Profile the Specific Cellular Targets of Andrographolide, a Promising Anticancer Agent That Suppresses Tumor Metastasis

A Quantitative Chemical Proteomics Approach to Profile the Specific Cellular Targets of Andrographolide, a Promising Anticancer Agent That Suppresses Tumor Metastasis

Nanomaterial-based miniaturized extraction and preconcentration techniques coupled to matrix-assisted laser desorption/ionization mass spectrometry for assaying biomolecules

Recent progress in application of carbon nanomaterials in laser desorption/ionization mass spectrometry

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