Organic Mercury Solid Phase Chemoselective Capture for proteomic identification of S-nitrosated proteins and peptides

Nitric Oxide ◽  
2021 ◽  
Author(s):  
Paschalis-Thomas Doulias ◽  
Margarita Tenopoulou ◽  
Iordanis Zakopoulos ◽  
Harry Ischiropoulos
Keyword(s):  
Solid Phase ◽  
Organic Mercury ◽  
Proteomic Identification ◽  
Proteins And Peptides

scholarly journals Sensitive Mercury Speciation by Reversed-Phase Column High-Performance Liquid Chromatography with UV-Visible Detection After Solid-Phase Extraction Using 6-Mercaptopurine and Dithizone

2008 ◽  
Vol 91 (6) ◽  
pp. 1453-1458 ◽  
Author(s):  
Hamid Hashemi-Moghaddam ◽  
Mohamad Saber-Tehrani
Keyword(s):  
High Performance ◽  
Solid Phase ◽  
Reversed Phase ◽  
Phase Extraction ◽  
Organic Mercury ◽  
Tuna Fish ◽  
Rp Hplc ◽  
Uv Visible ◽  
Phase Column ◽  
Reversed Phase Column

Abstract A highly selective and sensitive method was developed for preconcentration of inorganic and organic mercury compounds followed by reversed-phase column high-performance liquid chromatography (RP-HPLC) with UV-visible detection. The method was based on the reaction of mercury with 6-mercaptopurine and solid-phase extraction (SPE) of the complex on an octadecylsilane (C18) cartridge. The complex was then treated with ammoniacal dithizone solution, and the complexes of inorganic and organic mercury with dithizone were eluted by methanol. The speciation analysis of methylmercury (MeHg), phenylmercury (PhHg), and inorganic Hg (II) was carried out by RP-HPLC. Some experimental variables that influence the SPE and derivatization, such as pH, chelating and derivatizing agent concentration, and surfactant addition, were investigated. The calibration graphs of MeHg, PhHg, and Hg (II) were linear [correlation coefficient (r) > 0.999] from the detection limits (0.12, 0.16, and 0.14 ng) to 8.5, 6.0, and 6.7 ng Hg, respectively. By applying the SPE procedure, a 100-fold concentration of the sample was obtained. The procedure was applied to sea water and tuna fish samples. The method's accuracy was investigated by using tuna fish certified reference material BCR 464 and by spiking the samples with different amounts of MeHg, PhHg, and Hg (II). The average recoveries of MeHg, PhHg, and Hg (II) from spiked samples (0.12.0 g/L Hg) were 96 4, 98 3, and 104 4, respectively.

Development of Membrane Supports for the Solid-phase Sequence Analysis of Proteins and Peptides

1989 ◽  
pp. 69-78 ◽  
Author(s):  
James M. Coull ◽  
James D. Dixon ◽  
Richard A. Laursen ◽  
Hubert Köster ◽  
Darryl Pappin
Keyword(s):  
Sequence Analysis ◽  
Solid Phase ◽  
Phase Sequence ◽  
Proteins And Peptides

An enzymatic solid-phase method for trace iodination of proteins and peptides with 125iodine

1975 ◽  
Vol 67 (1) ◽  
pp. 1-10 ◽  
Author(s):  
S.-L. Karonen ◽  
P. Mörsky ◽  
M. Siren ◽  
U. Seuderling
Keyword(s):  
Solid Phase ◽  
Phase Method ◽  
Solid Phase Method ◽  
Proteins And Peptides

Utility of the gas-phase sequencer for both liquid- and solid-phase degradation of proteins and peptides at low picomole levels

1984 ◽  
Vol 140 (2) ◽  
pp. 553-566 ◽  
Author(s):  
James E. Strickler ◽  
Michael W. Hunkapiller ◽  
Kenneth J. Wilson
Keyword(s):  
Gas Phase ◽  
Solid Phase ◽  
Proteins And Peptides

scholarly journals Analysis of Albumin-Associated Peptides and Proteins from Ovarian Cancer Patients

2005 ◽  
Vol 51 (10) ◽  
pp. 1933-1945 ◽  
Author(s):  
Mark S Lowenthal ◽  
Arpita I Mehta ◽  
Kristina Frogale ◽  
Russell W Bandle ◽  
Robyn P Araujo ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Molecular Weight ◽  
Western Blotting ◽  
Cancer Susceptibility ◽  
Solid Phase ◽  
Reversed Phase ◽  
Low Molecular Weight ◽  
Affinity Capture ◽  
Proteins And Peptides

Abstract Background: Albumin binds low–molecular-weight molecules, including proteins and peptides, which then acquire its longer half-life, thereby protecting the bound species from kidney clearance. We developed an experimental method to isolate albumin in its native state and to then identify [mass spectrometry (MS) sequencing] the corresponding bound low–molecular-weight molecules. We used this method to analyze pooled sera from a human disease study set (high-risk persons without cancer, n= 40; stage I ovarian cancer, n = 30; stage III ovarian cancer, n = 40) to demonstrate the feasibility of this approach as a discovery method. Methods: Albumin was isolated by solid-phase affinity capture under native binding and washing conditions. Captured albumin-associated proteins and peptides were separated by gel electrophoresis and subjected to iterative MS sequencing by microcapillary reversed-phase tandem MS. Selected albumin-bound protein fragments were confirmed in human sera by Western blotting and immunocompetition. Results: In total, 1208 individual protein sequences were predicted from all 3 pools. The predicted sequences were largely fragments derived from proteins with diverse biological functions. More than one third of these fragments were identified by multiple peptide sequences, and more than one half of the identified species were in vivo cleavage products of parent proteins. An estimated 700 serum peptides or proteins were predicted that had not been reported in previous serum databases. Several proteolytic fragments of larger molecules that may be cancer-related were confirmed immunologically in blood by Western blotting and peptide immunocompetition. BRCA2, a 390-kDa low-abundance nuclear protein linked to cancer susceptibility, was represented in sera as a series of specific fragments bound to albumin. Conclusion: Carrier-protein harvesting provides a rich source of candidate peptides and proteins with potential diverse tissue and cellular origins that may reflect important disease-related information.

scholarly journals The promising use of nano-molecular imprinted templates for improved SARS-CoV-2 detection, drug delivery and research

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Alaa F. Nahhas ◽  
Thomas J. Webster
Keyword(s):  
Drug Delivery ◽  
Binding Sites ◽  
Solid Phase ◽  
Soft Materials ◽  
Disease Diagnosis ◽  
Antigenic Determinants ◽  
Extraction Separation ◽  
Therapeutic Tools ◽  
Proteins And Peptides ◽  
Complementary Binding

AbstractMolecular imprinting (MI) is a technique that creates a template of a molecule for improving complementary binding sites in terms of size and shape to a peptide, protein, bacteria, mammalian cell, or virus on soft materials (such as polymers, hydrogels, or self-assembled materials). MI has been widely investigated for over 90 years in various industries but is now focused on improved tissue engineering, regenerative medicine, drug delivery, sensors, diagnostics, therapeutics and other medical applications. Molecular targets that have been studied so far in MI include those for the major antigenic determinants of microorganisms (like bacteria or viruses) leading to innovations in disease diagnosis via solid-phase extraction separation and biomimetic sensors. As such, although not widely investigated yet, MI demonstrates much promise for improving the detection of and treatment for the current Coronavirus Disease of 2019 (COVID-2019) pandemic as well as future pandemics. In this manner, this review will introduce the numerous applications of MI polymers, particularly using proteins and peptides, and how these MI polymers can be used as improved diagnostic and therapeutic tools for COVID-19. Graphic Abstract

scholarly journals Evaluation of Sample Preparation Strategies for Human Milk and Plasma Proteomics

Molecules ◽  
2021 ◽  
Vol 26 (22) ◽  
pp. 6816
Author(s):  
Sanja Milkovska-Stamenova ◽  
Michele Wölk ◽  
Ralf Hoffmann
Keyword(s):  
Sample Preparation ◽  
Human Milk ◽  
Biological Sample ◽  
Solid Phase ◽  
Protein Precipitation ◽  
Efficient Procedure ◽  
Identification Rate ◽  
Advantages And Disadvantages ◽  
Peak Areas ◽  
Proteins And Peptides

Sample preparation is the most critical step in proteomics as it directly affects the subset of proteins and peptides that can be reliably identified and quantified. Although a variety of efficient and reproducible sample preparation strategies have been developed, their applicability and efficacy depends much on the biological sample. Here, three approaches were evaluated for the human milk and plasma proteomes. Protein extracts were digested either in an ultrafiltration unit (filter-aided sample preparation, FASP) or in-solution (ISD). ISD samples were desalted by solid-phase extraction prior to nRPC-ESI-MS/MS. Additionally, milk and plasma samples were directly digested by FASP without prior protein precipitation. Each strategy provided inherent advantages and disadvantages for milk and plasma. FASP appeared to be the most time efficient procedure with a low miscleavage rate when used for a biological sample aliquot, but quantitation was less reproducible. A prior protein precipitation step improved the quantitation by FASP due to significantly higher peak areas for plasma and a much better reproducibility for milk. Moreover, the miscleavage rate for milk, the identification rate for plasma, and the carbamidomethylation efficiency were improved. In contrast, ISD of both milk and plasma resulted in higher miscleavage rates and is therefore less suitable for targeted proteomics.

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