MICROCHIMICA ACTA
·
2019
·Vol. 187
·期 1
·1-12
·
IF 5.479
Zheng Haijiao,Jia Qiong
A hybrid monolith was prepared from perovskite and cucurbit[6]uril [poly(hydroxyethyl methacrylate-pentaerythritol triacrylate) monolith] for the enrichment of phosphopeptides. By coupling with mass spectrometry, three goals were simultaneously realized, viz. (a) selective enrichment of phosphopeptides from non-phosphopeptides, (b) identification of mono- and multi-phosphopeptides, and (c) recognition of tyrosine phosphopeptides. The perovskite introduced into the monolith warrants high selectivity for phosphopeptides even at a high (10,000:1) ratio of non-phosphopeptides to phosphopeptides, and and enables identification of eight mono- and multi-phosphopeptides from standard β-casein tryptic digests. Tyrosine phosphopeptides were specifically detected via the recognition capability of cucurbit[6]uril integrated into the monolith. The method has remarkably specific enrichment capacity for phosphopeptides from samples including human serum, nonfat milk, and human acute myelocytic leukemia cell lysate.
Schematic representation of a monolith integrated with perovskite and cucurbit[6]uril. The monolithic column was coupled with mass spectrometry and applied to the enrichment of phosphopeptides. The method has remarkably specific enrichment capacity for phosphopeptides from complex biological samples.
研究领域:
分析化学
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生物化学
·
蛋白质组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
·
2025
·
IF 12.5
Rong Li,Yan Wang,Dandan Zhou,Ruizhi Tang,Yangyang Bian,Shujuan Ma,Xingtai Qiu,Zichao Tang,Junjie Ou,Yehua Shen
Preferential enrichment of phosphopeptides with different numbers of phosphorylation sites is crucial to investigate the modulation of signal transduction pathways of a graded protein kinase or phosphatase signal. Here, zirconium phosphonates chitosan monoliths (ZrPCMs) with varying compositions were constructed through a salt-forming reaction and bidirectional freezing strategy. These composite monoliths exhibited differential coordination abilities for mono- and multi-phosphopeptides without the need for centrifugation or a magnetic field during enrichment process. By the utilization of different ZrPCMs and control of loading amount of sample, selective enrichment of mono- or multi-phosphopeptides was realized. At high loading amounts, excess phosphate groups on ZrPCM-1 inhibited mono-phosphopeptides with chelating with Zr (IV), resulting in the preferential enrichment of multi-phosphopeptides. In contrast, ZrPCM-2 enabled the enrichment of global phosphopeptides due to its balanced phosphate group and Zr(IV) loading, avoiding selective preferences under extreme conditions. At low loading amounts, the availability of sufficient Zr (IV) on ZrPCM-3 facilitated the preferential enrichment of mono-phosphopeptides due to their lower steric hindrance and more stable binding, while the adsorption ability for multi-phosphopeptides was reduced owing to local charge accumulation effects and electrostatic repulsion. These results were verified not only in standard protein but also in complex biological samples involving nonfat milk and mouse liver. Consequently, this present work developed promising ZrPCMs materials and isolation strategy of mono-, multi-, or global phosphopeptides for phosphoproteome research.
研究领域:
生物化学
·
分析化学
·
蛋白质组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2016
·
IF 4.035
Junyong Dai,Miaode Wang,Hailong Liu
It is urgent to selectively enrich trace phosphorylated proteins or peptides from complex biological sample for mass spectrometry (MS)-based phosphoproteomics. In this study, a novel immobilized metal affinity chromatography (IMAC) material with DOTA(1,4,7,10-tetraazacyclododecane N, N′, N′′, N′′′-tetra-acetic acid) coated on the surface of TiO 2 nanomaterials and functionalized with the metal ion Zr 4+ (TiO 2 @DOTA-Zr) has been synthesized successfully to improve the enrichment specificity and sensitivity for phosphopeptides. TiO 2 nanomaterials were chosen as a support matrix, which will offer a high specific surface area to enhance the amounts of immobilized Zr 4+ ions and will provide an additional selectivity for the special enrichment of phosphopeptides. Also, the introduction of DOTA will endow a stronger binding interaction between the Zr 4+ ion and DOTA to reduce the loss of the metal ion Zr 4+ during sample loading and washing steps. The novel TiO 2 @DOTA-Zr nanohybrids exhibited higher sensitivity and selectivity for phosphopeptides from the standard protein digests, the tryptic digests of nonfat milk and human serum, showing a great selective enrichment capability toward the complicated biological samples.
研究领域:
生物化学
·
蛋白质组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2014
·
IF 4.258
Xiao-Shui Li,Xi Chen,Huan Sun,Bi-Feng Yuan,Yu-Qi Feng
Selective and effective enrichment of phosphopeptides from complex samples is essential in phosphoproteome study by mass spectrometry (MS). In this work, we compared perovskites (MgTiO 3 , CaTiO 3 , SrTiO 3 , BaTiO 3 and CaZrO 3 ) with metal oxides (ZrO 2 and TiO 2 ) in their capability for the selective enrichment of phosphopeptides. It was found here that perovskites exhibited higher selectivity towards phosphopeptides than commonly used ZrO 2 and TiO 2 , even though they all have high affinity to phosphopeptides. As for perovskites, CaTiO 3 exhibited better selectivity for enrichment of phosphopeptides than SrTiO 3 , MgTiO 3 , BaTiO 3 and CaZrO 3 , which might be ascribed to their crystal structures and electrophilic abilities. Moreover, to further confirm the performance of CaTiO 3 , CaTiO 3 and TiO 2 were applied to the enrichment of phosphopeptides from tryptic digest of proteins of human Jurkat-T cell lysate, respectively. The results showed CaTiO 3 has much higher selectivity than TiO 2 in the enrichment of phosphopeptides from the complex biological sample. Taken together, here we show that CaTiO 3 is an excellent material for the highly selective enrichment of phosphopeptides and it could be potentially used in the large-scale phosphoproteome study.
研究领域:
蛋白质组学
·
分析化学
·
生物医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Feifei Xu,Danyi Shang,Congcong Zhu,Guangzhu Du,Jingchen Shi,Xuefang Dong,Xiuling Li,Xinmiao Liang
Phosphopeptide enrichment methods based on commercial TiO2 suffer from difficulties in regulating intermolecular interactions, resulting in low coverage rate and the loss of information on multiphosphorylation sites, thereby limiting comprehensive phosphoproteomic analysis. In this work, MXene Ti3C2Tx was incorporated into the design of enrichment materials, with its surface structure functionalized and regulated to address the low elution efficiency of TiO2 for multiphosphorylated peptides. Upon oxidation treatment, the Ti3C2Tx material formed numerous uniformly distributed TiO2 nanoparticles on the surface of Ti3C2Tx-O, providing abundant affinity sites (Ti–O) for selective phosphopeptide enrichment. The polycrystalline structure and rich oxygen vacancies of the material effectively regulated its binding affinity with phosphate groups, achieving simultaneous high-efficiency enrichment of both monophosphorylated and multiphosphorylated peptides. Its performance was significantly superior to that of commercial TiO2 and IMAC materials. This study presents great promise for the practical application of comprehensive phosphoproteomic analysis in the future and broadens the application of MXene in the biological field.
研究领域:
分析化学
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生物化学
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Functional Biomaterials
·
2024
·Vol. 15
·期 7
·185
·
IF 5
Yu Wu,Sen Xu,Fengjuan Ding,Weibing Zhang,Haiyan Liu
In this work, a new type of FE-1 COF material is prepared by a reversible imine condensation reaction with diaminoferrocene and diaminodiformaldehyde as materials. The material is connected by imine bonds to form a COF skeleton, and the presence of plenty of nitrogen-containing groups gives the material good hydrophilicity; the presence of metal Fe ions provides the material application potential in the enrichment of phosphopeptides. According to the different binding abilities of N-glycopeptide and phosphopeptide on FE-1 COF, it can simultaneously enrich N-glycopeptide and phosphopeptide through different elution conditions to realize its controllable and selective enrichment. Using the above characteristics, 18 phosphopeptides were detected from α-casein hydrolysate, 8 phosphopeptides were detected from β-casein hydrolysate and 21 glycopeptides were detected from IgG hydrolysate. Finally, the gradual elution strategy was used; 16 phosphopeptides and 19 glycopeptides were detected from the α-casein hydrolysate and IgG hydrolysate. The corresponding glycopeptides and phosphopeptides were identified from the human serum. It proves that the FE-1 COF material has a good enrichment effect on phosphopeptides and glycopeptides.
研究领域:
分析化学
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蛋白质组学
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生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2024
·
IF 2.8
FangKun Ren,JunYong Dai,JingYi Zhang,YanFei Luan,Fan Yang,Jian Shen,HaiLong Liu,JiaHong Zhou
The specific enrichment of multi-phosphopeptides in the presence of non-phosphopeptides and mono-phosphopeptides was still a challenge for phosphoproteomics research. Most of these enrichment materials relied on Zn, Ti, Sn, and other rare precious metals as the bonding center to enrich multi-phosphopeptides while ignoring the use of common metal elements. The addition of rare metals increased the cost of the experiment, which was not conducive to their large-scale application in biomedical proteomics laboratories. In addition, multiple high-speed centrifugation steps also resulted in the loss of low-abundance multi-phosphopeptides in the treatment procedure of biological samples. This study proposed the use of calcium, a common element, as the central bonding agent for synthesizing magnetic calcium phosphate materials (designated as CaP-Fe 3 O 4 ). These materials aim to capture multi-phosphopeptides and identifying phosphorylation sites. The current results demonstrate that CaP-Fe 3 O 4 exhibited excellent selection specificity, high sensitivity, and stability in the enrichment of multi-phosphopeptides and the identification of phosphorylation sites. Additionally, the introduction of magnetic separation not only reduced the time required for multi-phosphopeptides enrichment but also prevented the loss of these peptides during high-speed centrifugation. These findings contribute to the widespread application and advancement of phosphoproteomics research.
研究领域:
生物化学
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蛋白质组学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2024
·Vol. 16
·期 5
·695-703
·
IF 2.7
Jiayi Wang,Xiaoya Zhang,Yinghua Yan,Rongrong Xuan
The comprehensive investigation of protein phosphorylation and glycosylation aids in the discovery of novel biomarkers as well as the understanding of the pathophysiology of illness. In this work, a nitrogen/titanium-rich porous organic polymer was developed by copolymerizing carbohydrazide (CH) and 2,3-dihydroxyterephthalaldehyde (2,3-Dha) and modifying with Ti4+ (CH–Dha–Ti4+). The adequate nitrogen contributes to the enrichment of glycopeptides via HILIC, while titanium benefits from capturing phosphopeptides through IMAC. The proposed method exhibits excellent selectivity (1 : 1000, both for glycopeptides and phosphopeptides), LOD (for glycopeptides: 0.05 fmol μL−1, for phosphopeptides: 0.2 fmol), loading capacity (for glycopeptides: 100 mg g−1, for phosphopeptides: 125 mg g−1) and size-exclusion effect (1 : 10 000, both for glycopeptides and phosphopeptides). Furthermore, CH–Dha–Ti4+ was applied to capture glycopeptides and phosphopeptides from human serum; 205 glycopeptides and 45 phosphopeptides were detected in the serum of normal controls; and 294 glycopeptides and 63 phosphopeptides were found in the serum of uremia patients after being analyzed by nano LC-MS/MS. The discovered glycopeptides and phosphopeptides were involved in several molecular biological processes and activities, according to a gene ontology study.
研究领域:
蛋白质组学
·
生物化学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Ruizhi Tang,Yang Yu,Jing Dong,Yating Yao,Shujuan Ma,Junjie Ou,Mingliang Ye
In bottom-up strategy, specific enrichment of glycopeptides and phosphopeptides from complicated biological samples is a prerequisite for efficient identifying glycosylation and phosphorylation by mass spectrometry . Although there were a plethora of materials used as either hydrophilic interaction liquid chromatography (HILIC) or immobilized metal affinity chromatography (IMAC) adsorbents, even several bifunctional materials for simultaneous enrichment of glycopeptides and phosphopeptides, most of them are not easily commercialized as many other well-performing adsorbents due to the complicated preparation process. In our case, a one-step modification strategy was developed to prepare bifunctional adsorbents for HILIC and IMAC, employing O -phospho- l -serine as the modifier and poly(GMA- co -EDMA) microspheres, a kind of macroporous adsorption resin (MAR) with epoxy groups , as the matrix. The MARs were directly modified with O -phospho- l -serine under facile condition for HILIC strategy and further chelated with Ti 4+ for IMAC strategy. A total of 522 unique N -glycopeptides and 442 unique N -glycosylation sites mapped to 275 N -glycoproteins was identified from HeLa cell proteins, showing excellent enrichment efficiency in HILIC. Additionally, 3141 unique phosphopeptides were unambiguously identified from 200 μg of digest of HeLa cell proteins, demonstrating great enrichment efficiency in IMAC. Moreover, these materials have been successfully applied in the analysis of multiple biological samples including human serum and milk, demonstrating their feasibility for real sample applications and potential business value.
研究领域:
蛋白质组学
·
生物化学
·
分析化学
✓ 已验证
DOI
高影响因子
TALANTA
·
2015
·
IF 3.545
Haizhu Lin,Chunhui Deng
In this work, we first designed and synthesized an IMAC material with Hf 4+ immobilized on polydopamine-coated magnetic graphene and applied it to phosphopeptides enrichment successfully. The newly prepared material gathered the advantages of large specific surface area of graphene, good hydrophilicity and biocompatibility of polydopamine and superparamagnetism of Fe 3 O 4 . We investigated the performance of Hf 4+ -immobilized polydopamine-coated magnetic graphene (denoted as magG@PDA-Hf 4+ ) in phosphopeptides enrichment and the results showed high selectivity and sensitivity of the new material. Finally, we successfully applied magG@PDA-Hf 4+ to phosphopeptides enrichment from non-fat milk digests and human serum, further demonstrating excellent performance of this new material in phosphopeptides enrichment.
研究领域:
生物化学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 12
·1-8
·
IF 5.705
Lin Haizhu,Chen Hemei,Shao Xi,Deng Chunhui
A zirconium(IV)-based metal organic framework (Zr-MOF) was deposited on polydopamine-coated silica microspheres to form microspheres of type SiO 2 @PDA@Zr-MOF. These were packed into capillary columns for enrichment of phosphopeptides. The column was off-line coupled to both matrix-assisted laser desorption/ionization time of flight mass spectrometry and LC-ESI-MS/MS. The method has a detection limit as low as 4 fmol of β-casein digest and a selectivity as high as 1:1000 (molar ratio of β-casein and BSA digest). It was applied to the analysis of human saliva. In total, 240 endogenous phosphopeptides were identified in only 25 μL human saliva.
A zirconium-based metal organic framework (Zr-MOF) was modified outside of polydopamine-coated silica microspheres to form microspheres named SiO 2 @PDA@Zr-MOF. Then they were packed in capillary columns for selective enrichment of phosphopeptides via interaction between Zr-O clusters and phosphate groups. The pre-concentration resulted in a better detection of phosphopeptides by mass spectrometry. Tris: Tris(hydroxymethyl)aminomethane; DMF: Dimethyl Formamide; Zr-MOF: Zirconium(IV)-organic framework; MOAC: Metal oxide affinity chromatography.
研究领域:
分析化学
·
生物医学
✓ 已验证
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Jie Li,Junlong Wang,Jiajun Cui,Fei Gao,Hongying Guo,Jiyou Yang,Mengren Li,Zhan Ma,Zijun Wang,Liuping Zhang,Weiwei Huan
Natural plants with fascinating structures give inspirations to fabricate advanced affinity chromatography materials for glycoproteome and phosphoproteome analyses. In this study, inspired by the long-range ordered structure and rapid water transportation capability of lotus stem, a biomimetic hydrophilic material (PE/HAVP-Ti 4+ ) with parallel ordered channels is developed and applied to selectively enrich low-abundant glycopeptides and phosphopeptides in biological samples. The biomimetic PE/HAVP-Ti 4+ has honeycomb porous structure, robust mechanical property, excellent hydrophilicity, and abundant affinity sites. It demonstrates the excellent selectivity (molar ratio human IgG: BSA=1:1000, β-casein: BSA=1:2500), high sensitivity (0.2 fmol μL –1 for glycopeptides, 0.01 fmol μL –1 for phosphopeptides), satisfactory recovery rate (90.7 % for glycopeptides, 91.1 % for phosphopeptides), and good recyclability (at least 10 cycles). Moreover, after enrichment treatment by PE/HAVP-Ti 4+ , a total of 411 N - glycopeptides assigned to 212 N-glycoproteins and 63 phosphopeptides associated with 41 phosphoproteins in human serum are identified by mass spectrometry. The PE/HAVP-Ti 4+ has significant potential in applications of glycopeptide and phosphopeptide enrichment of real complex biological samples for the simultaneous study of glycoproteome and phosphoproteome.
研究领域:
生物化学
·
分析化学
·
蛋白质组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）