Although bottom-up proteomics using tryptic digests is widely used to locate post-translational modifications (PTM) in proteins, there are cases where the protein has several potential modification sites within a tryptic fragment and MS(2) strategies fail to pinpoint the location. We report here a method using two proteolytic enzymes, trypsin and pepsin, in combination followed by tandem mass spectrometric analysis to provide fragments that allow one to locate the modification sites. We used this strategy to find a glycosylation site on bovine trypsin expressed in maize (TrypZean). Several glycans are present, and all are attached to a nonconsensus N-glycosylation site on the protein.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3005374 | PMC |
| http://dx.doi.org/10.1021/ac1020722 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
The Enzyme Effect: Broadening the Horizon of MS Optimization to Nontryptic Digestion in Proteomics.
J Am Soc Mass Spectrom
January 2025
MS Proteomics Research Group, HUN-REN Research Centre for Natural Sciences, Magyar Tudósok körútja 2, H-1117 Budapest, Hungary.
In recent years, alternative enzymes with varied specificities have gained importance in MS-based bottom-up proteomics, offering orthogonal information about biological samples and advantages in certain applications. However, most mass spectrometric workflows are optimized for tryptic digests. This raises the questions of whether enzyme specificity impacts mass spectrometry and if current methods for nontryptic digests are suboptimal.
View Article and Find Full Text PDFDesalting of oligonucleotides through precipitation for mass spectrometric analysis.
Nucleosides Nucleotides Nucleic Acids
January 2025
Department of Chemistry and Centre for Biotechnology, Brock University, St. Catharines, ON, Canada.
Contamination of sodium ions in oligonucleotides often causes issues in mass spectrometric analysis. This study investigated the efficiency of the combination of ammonium acetate and alcohol in desalting oligonucleotides. It was found that oligonucleotide samples containing up to 4 M NaCl can be effectively desalted through precipitation with ethanol or isopropanol in the presence of 1 or 5 M ammonium acetate.
View Article and Find Full Text PDFSacubitril/valsartan reduces proteasome activation and cardiomyocyte area in an experimental mouse model of hypertrophy.
J Mol Cell Cardiol Plus
March 2024
Institute of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Martinistr. 52, 20246 Hamburg, Germany.
Sacubitril/valsartan (Sac/Val) belongs to the group of angiotensin receptor-neprilysin inhibitors and has been used for the treatment of heart failure (HF) for several years. The mechanisms that mediate the beneficial effects of Sac/Val are not yet fully understood. In this study we investigated whether Sac/Val influences the two proteolytic systems, the ubiquitin-proteasome system (UPS) and the autophagy-lysosomal pathway (ALP), in a mouse model of pressure overload induced by transverse aortic constriction (TAC) and in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) treated with endothelin-1 (ET1) serving as a human cellular model of hypertrophy.
View Article and Find Full Text PDFYiqi Huayu Jiedu formula inhibits JAK2/STAT3-mediated partial EMT in treating chronic atrophic gastritis.
Phytomedicine
December 2024
State Key Laboratory of Traditional Chinese Medicine Syndrome/Department of Gastroenterology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangdong Provincial Hospital of Chinese Medicine, Guangdong Provincial Academy of Chinese Medical Sciences/State Key Laboratory of Dampness Syndrome of Chinese Medicine, Guangzhou, China; Guangdong Provincial Key laboratory of Chinese Medicine for Prevention and Treatment of Refractory Chronic Diseases, Guangzhou, China. Electronic address:
Background: Chronic atrophic gastritis (CAG) is a precursor to gastric cancer, a leading cause of cancer-related deaths worldwide. Despite current therapeutic strategies, preventing the transition from gastritis to cancer remains a challenge. Traditional Chinese Medicine (TCM), particularly the Yiqi-Huayu-Jiedu (YQHYJD) formula, have exhibited promising results in CAG management.
View Article and Find Full Text PDFMALDI-TOF mass spectrometric immunoassay of multiple tumor biomarkers for non-small cell lung cancer screening.
Talanta
January 2025
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China. Electronic address:
Cancer biomarkers have been facing some issues such as poor accuracy and low sensitivity in the early diagnosis of tumors. Utilizing biotin-labelled peptide as a mass tag (MT), this work proposes a high-throughput biosensing strategy for matrix-assisted laser desorption/ionization-time of flight mass spectrometric (MALDI-TOF-MS) immunoassay of multiple lung cancer biomarkers. Due to little required dosage, satisfied stability, high sensitivity and accuracy, this method can achieve off-site centralized signal detection after on-site sample incubation.
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!