scholarly journals Binder-Free TiO2 Monolith-Packed Pipette Tips for the Enrichment of Phosphorylated Peptides

2016 ◽  
Vol 69 (12) ◽  
pp. 1396 ◽  
Author(s):  
Chang Lei ◽  
Liang Zhou ◽  
Chun Xu ◽  
Xiaoran Sun ◽  
Amanda Nouwens ◽  
...  
Keyword(s):  
Detection Limit ◽  
Pure Tio2 ◽  
Phosphorylated Peptides ◽  
Binder Free ◽  
Peptide Enrichment ◽  
Pipette Tip ◽  
Phosphorylated Peptide ◽  
Packing Method ◽  
Enrichment Efficiency

A macroporous TiO2 monolith-entrapped pipette-tip was developed through a binder-free packing method for convenient phosphorylated peptide enrichment. A detection limit of 1 ng mL–1 for phosphorylated peptide is achieved, showing a better enrichment efficiency compared with the commercial pure TiO2-embedded NuTip.

scholarly journals Skimmer CID‒ion trap mass spectrometry of phosphotyrosine‒containing peptides

2004 ◽  
Vol 18 (3) ◽  
pp. 441-451
Author(s):  
Melissa D. Zolodz ◽  
Karl V. Wood
Keyword(s):  
Mass Spectrometry ◽  
Ion Trap ◽  
Phosphorylation Site ◽  
Ion Trap Mass Spectrometry ◽  
Cellular Functions ◽  
Factors Affecting ◽  
Parent Protein ◽  
Phosphorylated Peptides ◽  
Phosphorylated Peptide ◽  
Analytical Tools

Proteomic analysis is becoming a popular field in science. Analysis of protein modifications is useful in deciphering cellular functions and errors in pathways that can result in disease. There has been increased interest in the phosphotyrosine proteome. Due to the difficulty in finding the location of the tyrosine phosphorylation site in the tyrosine phosphorylated peptide or even to verify that the parent protein is a phosphotyrosyl‒protein, new analytical tools are being developed. The phosphotyrosine immonium ion can be produced via skimmer CID for detection via ion trap mass spectrometry and is a useful marker for the indication of the presence of a phosphotyrosine residue. Skimmer CID analysis can also be used to differentiate phosphotyrosine‒containing peptides from other phosphorylated peptides. In this study, phosphotyrosine‒containing peptides were analyzed by skimmer CID in an ion trap mass spectrometer. The factors affecting the signal abundance of the phosphotyrosine immonium ion were investigated.

scholarly journals A Two-Dimensional Metallacycle Cross-Linked Switchable Polymer for Fast and Highly Efficient Phosphorylated Peptide Enrichment

2019 ◽  
Author(s):  
Li-Jun Chen ◽  
jun-long zhu ◽  
Fan-Fan Zhu ◽  
Jin Wen ◽  
Hai-Bo Yang ◽  
...  
Keyword(s):  
Detection Sensitivity ◽  
Supramolecular Polymer ◽  
Two Dimensional ◽  
Phosphopeptide Enrichment ◽  
Stimuli Responsive ◽  
Excellent Performance ◽  
High Enrichment ◽  
Peptide Enrichment ◽  
Phosphorylated Peptide ◽  
Positively Charged

The selective and efficient capture of phosphopeptides is critical for comprehensive and in-depth phosphoproteome analysis. However, this remains a significant challenge due to the inherently low abundance of these species in complex bio-samples. In this paper, we report a switchable two-dimensional (2D) supramolecular polymer that can serve as an ideal platform for the enrichment of phosphopeptides. A positively charged metallacycle incorporated into the polymer endows the material with a high affinity for phosphopeptides. Importantly, the stimuli-responsive nature of the polymer facilitates switchable binding affinity of phosphopeptides, resulting in its excellent performance in phosphopeptide enrichment and separation from model proteins. The polymer has a high enrichment capacity (165 mg/g) and detection sensitivity (2 fmol), high enrichment recovery (88%), excellent specificity, and rapid enrichment and separation properties. Additionally, we have demonstrated the capture of phosphopeptides from the tryptic digest of real bio-samples illustrating the potential of this polymeric material in phosphoproteomic studies.

A Cu2O/Cu/carbon cloth as a binder-free electrode for non-enzymatic glucose sensors with high performance

2020 ◽  
Vol 44 (5) ◽  
pp. 1993-2000 ◽  
Author(s):  
Haoze Zhang ◽  
Yawei Yu ◽  
Xiaodong Shen ◽  
Xiulan Hu
Keyword(s):  
Detection Limit ◽  
Electrochemical Deposition ◽  
High Performance ◽  
Glucose Sensors ◽  
Linear Range ◽  
Carbon Cloth ◽  
Binder Free

An electrode prepared via potentiostatic electrochemical deposition exhibits a 60 nM detection limit and a 1 linear range of 1 to 1555 μM.

scholarly journals K depletion increases protein tyrosine kinase-mediated phosphorylation of ROMK

2002 ◽  
Vol 283 (4) ◽  
pp. F671-F677 ◽  
Author(s):  
Dao-Hong Lin ◽  
Hyacinth Sterling ◽  
Kenneth M. Lerea ◽  
Paul Welling ◽  
Lianhong Jin ◽  
...  
Keyword(s):  
Protein Tyrosine Kinase ◽  
Signal Transduction Pathway ◽  
Tyrosine Residue ◽  
Deficient Diet ◽  
High K ◽  
Phosphorylated Peptides ◽  
K Secretion ◽  
Phosphorylated Peptide ◽  
Phosphopeptide Mapping

We purified His-tagged ROMK1 and carried out in vitro phosphorylation assays with 32P-radiolabeled ATP to determine whether ROMK1 protein is a substrate for PTK. Addition of active c-Src and [32P]ATP to the purified ROMK1 protein resulted in the phosphorylation of the ROMK1 protein. However, c-Src did not phosphorylate R1Y337A in which tyrosine residue 337 was mutated to alanine. Furthermore, phosphopeptide mapping identified two phosphopeptides from the trypsin-digested ROMK1 protein. In contrast, no phosphorylated peptide has been found in the trypsin-digested R1Y337A protein. This suggested that two phosphorylated peptides might contain the same tyrosine residue. Also, addition of c-Src and [32P]ATP phosphorylated the synthesized peptide corresponding to amino acid sequence 333–362 of the COOH terminus of ROMK1. We then examined the effect of dietary K intake on the tyrosine-phosphorylated ROMK level. Although the ROMK channels pulled down by immunoprecipitation with ROMK antibody were the same from rats on a K-deficient diet or on a high-K diet, more ROMK channels were phosphorylated by PTK in rats on a K-deficient diet than those on a high-K diet. We conclude that ROMK1 can be phosphorylated by PTK and that tyrosine residue 337 is the key site for the phosphorylation. Also, the tyrosine phosphorylation of ROMK is modulated by dietary K intake. This strongly suggests that PTK is an important member of the aldosterone-independent signal transduction pathway for regulating renal K secretion.

scholarly journals Examining ubiquitinated peptide enrichment efficiency through an epitope labeled protein

2016 ◽  
Vol 512 ◽  
pp. 114-119
Author(s):  
J. Parker ◽  
Y. Oh ◽  
Y. Moazami ◽  
J.G. Pierce ◽  
P.L. Loziuk ◽  
...  
Keyword(s):  
Peptide Enrichment ◽  
Enrichment Efficiency

scholarly journals Chromium-based metal organic framework for pipette tip micro-solid phase extraction: an effective approach for determination of methyl and propyl parabens in wastewater and shampoo samples

BMC Chemistry ◽  
2021 ◽  
Vol 15 (1) ◽  
Author(s):  
Massoud Kaykhaii ◽  
Sayyed Hossein Hashemi ◽  
Fariba Andarz ◽  
Amin Piri ◽  
Ghasem Sargazi ◽  
...  
Keyword(s):  
Detection Limit ◽  
Solid Phase Extraction ◽  
Solid Phase ◽  
Metal Organic Framework ◽  
Phase Extraction ◽  
Propyl Paraben ◽  
Metal Organic ◽  
Pipette Tip ◽  
Organic Framework

Abstract Background A chromium-based metal organic framework was synthesized and employed as an efficient sorbent for pipette tip micro-solid phase extraction and preconcentration of parabens from wastewater and shampoo samples up to sub-ppb level before their spectrophotometric analysis. Results Factors affecting preconcentration including volume and type of solvent, amount of sorbent, number of extraction, and volume and pH of samples were optimized employing one-variable-at-a-time and response surface methodology. Obtained analytical characteristics of the method proves its usefulness for analysis of real samples. Linear range of the method for parabens was 1.0–200.0 μg/L. Detection limit of the protocol was 0.24 µg/L for propyl paraben and 0.25 µg/L for methyl paraben. Reproducibility of the protocol defined as % RSD was better than 5.78%. Synthesized adsorbent can be re-used for at least 20 extractions. Conclusion The method showed a good detection limit and precision for determination of methyl- and propyl-paraben in wastewater and shampoo samples.

scholarly journals A Two-Dimensional Metallacycle Cross-Linked Switchable Polymer for Fast and Highly Efficient Phosphorylated Peptide Enrichment

2020 ◽  
Author(s):  
Li-Jun Chen ◽  
jun-long zhu ◽  
Fan-Fan Zhu ◽  
Xu-Qing Wang ◽  
Jin Wen ◽  
...  
Keyword(s):  
Detection Sensitivity ◽  
Supramolecular Polymer ◽  
Two Dimensional ◽  
Phosphopeptide Enrichment ◽  
Stimuli Responsive ◽  
Excellent Performance ◽  
High Enrichment ◽  
Peptide Enrichment ◽  
Phosphorylated Peptide ◽  
Positively Charged

<p>The selective and efficient capture of phosphopeptides is critical for comprehensive and in-depth phosphoproteome analysis. However, this remains a significant challenge due to the inherently low abundance of these species in complex bio-samples. In this paper, we report a switchable two-dimensional (2D) supramolecular polymer that can serve as an ideal platform for the enrichment of phosphopeptides. A positively charged metallacycle incorporated into the polymer endows the material with a high affinity for phosphopeptides. Importantly, the stimuli-responsive nature of the polymer facilitates switchable binding affinity of phosphopeptides, resulting in its excellent performance in phosphopeptide enrichment and separation from model proteins. The polymer has a high enrichment capacity (165 mg/g) and detection sensitivity (2 fmol), high enrichment recovery (88%), excellent specificity, and rapid enrichment and separation properties. Additionally, we have demonstrated the capture of phosphopeptides from the tryptic digest of real bio-samples illustrating the potential of this polymeric material in phosphoproteomic studies.</p>

scholarly journals A Two-Dimensional Metallacycle Cross-Linked Switchable Polymer for Fast and Highly Efficient Phosphorylated Peptide Enrichment

2020 ◽  
Author(s):  
Li-Jun Chen ◽  
jun-long zhu ◽  
Fan-Fan Zhu ◽  
Xu-Qing Wang ◽  
Jin Wen ◽  
...  
Keyword(s):  
Detection Sensitivity ◽  
Supramolecular Polymer ◽  
Two Dimensional ◽  
Phosphopeptide Enrichment ◽  
Stimuli Responsive ◽  
Excellent Performance ◽  
High Enrichment ◽  
Peptide Enrichment ◽  
Phosphorylated Peptide ◽  
Positively Charged

<p>The selective and efficient capture of phosphopeptides is critical for comprehensive and in-depth phosphoproteome analysis. However, this remains a significant challenge due to the inherently low abundance of these species in complex bio-samples. In this paper, we report a switchable two-dimensional (2D) supramolecular polymer that can serve as an ideal platform for the enrichment of phosphopeptides. A positively charged metallacycle incorporated into the polymer endows the material with a high affinity for phosphopeptides. Importantly, the stimuli-responsive nature of the polymer facilitates switchable binding affinity of phosphopeptides, resulting in its excellent performance in phosphopeptide enrichment and separation from model proteins. The polymer has a high enrichment capacity (165 mg/g) and detection sensitivity (2 fmol), high enrichment recovery (88%), excellent specificity, and rapid enrichment and separation properties. Additionally, we have demonstrated the capture of phosphopeptides from the tryptic digest of real bio-samples illustrating the potential of this polymeric material in phosphoproteomic studies.</p>

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