Conditions are presented for separating the major tryptic peptides of E.coli tryptophanyl-transfer RNA synthetase by reversed-phase liquid chromatography using a water-methanol gradient in the presence of 0.1% trifluoroacetic acid. Three of the peptides contain cysteine and are recovered in good yields if alkylated, but otherwise cannot be detected. A convenient post-digestion alkylation procedure is appropriate for use with small samples of protein which can be digested under reducing conditions. These results will be of interest for studies of the labeling of sulfhydryl groups in other proteins.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/0014-5793(85)80800-0 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Capillary Isoelectric Focusing of Proteins and Peptides Using an In-Line cIEF-ESI Interface with Improved MS Characteristics.
Anal Chem
January 2025
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, United States.
Intact protein analysis using mass spectrometry (MS) is an important technique to characterize and provide a comprehensive overview of protein complexity. It is also the basis of "top-down" approaches in proteomics to describe the proteoforms of single protein's post-translational modifications (PTMs). MS-based analysis of intact proteins benefits from high-resolution separations prior to electrospray ionization.
View Article and Find Full Text PDFUmbrella Sampling MD Simulations for Retention Prediction in Peptide Reversed-phase Liquid Chromatography.
Anal Chem
December 2024
Department of Chemistry, The University of Western Ontario, London, Ontario N6A 5B7, Canada.
Reversed-phase liquid chromatography (RPLC) is an essential tool for separating complex mixtures such as proteolytic digests in bottom-up proteomics. There is growing interest in methods that can predict the RPLC retention behavior of peptides and other analytes. Already, existing algorithms provide excellent performance based on empirical rules or large sets of RPLC training data.
View Article and Find Full Text PDFSimultaneous isotyping and semi-quantitation of anti-drug antibodies to an IgG1 biotherapeutic using hybrid LBA-LC-MS/MS.
Bioanalysis
December 2024
ICON Bioanalytical Laboratories, Assen, The Netherlands.
Background: Commonly, ligand-binding platforms are being used for immunogenicity assessment, but with the recent advent of liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) for protein quantification, this technology has become an alternative for the measurement of anti-drug antibodies (ADAs), when combined with an immunocapture step to extract them out of the biological sample.
Method: The monoclonal antibody adalimumab was immobilized on magnetic beads to isolate ADAs against this drug from serum samples. Multiple repetitions of immunopurification were used to minimize nonspecific binding and improve drug tolerance while maintaining sufficient recovery.
Mass Spectrometry-Based Workflow for the Identification and Quantification of Alternative and Canonical Proteins in Pancreatic Cancer Cells.
Cells
November 2024
Laboratoire de Recherche en Sciences Végétales (LRSV), CNRS/UT3/INPT, 31320 Auzeville-Tolosane, France.
The identification of small proteins and proteins produced from unannotated open reading frames (called alternative proteins or AltProts) has changed our vision of the proteome and has attracted more and more attention from the scientific community. Despite several studies investigating particular AltProts in diseases and demonstrating their importance in such context, we are still missing data on their expression and functions in many pathologies. Among these, pancreatic ductal adenocarcinoma (PDAC) is a particularly relevant case to study alternative proteins.
View Article and Find Full Text PDFHigh-Recovery Desalting Tip Columns for a Wide Variety of Peptides in Mass Spectrometry-Based Proteomics.
Anal Chem
December 2024
Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan.
In mass spectrometry-based proteomics, loss-minimized peptide purification techniques play a key role in improving sensitivity and coverage. We have developed a desalting tip column packed with thermoplastic polymer-coated chromatographic particles, named ChocoTip, to achieve high recoveries in peptide purification by pipet-tip-based LC with centrifugation (tipLC). ChocoTip identified more than twice as many peptides from 20 ng of tryptic peptides from Hela cell lysate compared to a typical StageTip packed with chromatographic particles entangled in a Teflon mesh in tipLC.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!