Research discoveries: glycoprotein glycosylation
New research papers from the Zaia laboratory have established novel strategies for analyzing glycans and glycopeptides.
Glycomics and glycoproteomics analyses by mass spectrometry require efficient front-end separation methods to enable deep characterization of heterogeneous glycoform populations. Chromatography methods a
re generally limited in their ability to resolve glycoforms using mobile phases that are compatible with online liquid chromatography-mass spectrometry (LC-MS). The adoption of capillary electrophoresis-mass spectrometry methods (CE-MS) for glycomics and glycoproteomics is limited by the lack of convenient interfaces for coupling the CE devices to mass spectrometers. Here, we describe the application of a microfluidics-based CE-MS system for analysis of released glycans, glycopeptides and monosaccharides. We demonstrate a single CE method for analysis three different modalities, thus contributing to comprehensive glycoproteomics analyses. In addition, we explored compatible sample derivatization methods. We used glycan TMT-labeling to improve electrophoretic migration and enable multiplexed quantitation by tandem MS. We used sialic acid linkage-specific derivatization methods to improve separation and the level of information obtained from a single analytical step. Capillary electrophoresis greatly improved glycoform separation for both released glycans and glycopeptides over that reported for chromatography modes more frequently employed for such analyses. Overall, the CE-MS method described here enables rapid setup and analysis of glycans and glycopeptides using mass spectrometry
To learn more:
- Khatri, K.; Klein, J. A.; Zaia, J. Use of an informed search space maximizes confidence of site-specific assignment of glycoprotein glycosylation. Anal Bioanal Chem 2017, 409, 607-618. Pubmed Link
- Khatri, K.; Klein, J. A.; Haserick, J.; Leon, D. R.; Costello, C. E.; McComb, M. E.; Zaia, J. Microfluidic capillary electrophoresis-mass spectrometry for analysis of monosaccharides, oligosaccharides and glycopeptides. Anal. Chem. 2017, 89, 6645-6655. Pubmed Link
Dahoud Breast Cancer Pilot Awards
Congratulations to several Biochemistry faculty who were recently awarded Dahoud Breast Cancer Pilot Awards.
Alla Grishok, PhD, Associate Professor of Biochemistry, Dafne Cardamone, PhD, Instructor, and Catherine Costello, PhD, Director of Center for Biomedical Mass Spectrometry, will study regulation of cancer-promoting Myc protein using a model metastatic breast cancer cell line. Myc binds DNA and activates a large network of genes that together transform normal cells into cancer cells. Myc activity is elevated in most human cancers and is especially relevant for Myc-driven triple (estrogen, progesterone and Her2) negative breast cancer. Dr. Grishok and colleagues will investigate new mechanisms that increase Myc protein activity: 1) adding specific sugar residues, and 2) protein truncation. New compounds that directly inhibit Myc or inhibit enzymes that activate Myc could be developed into new cancer therapies.
Xaralabos Varelas, PhD, Associate Professor of Biochemistry and Stefano Monti, PhD, Associate Professor of Medicine and Biostatistics, will study the causes of aggressive triple negative breast cancers. The team will determine how abnormal signaling networks drive gene expression changes that lead to aggressive breast cancers and then categorize subsets of aggressive breast cancers, thereby better targeting the most effective treatments based on the genes expressed in the tumor.
Cathy Costello receives prestigious mass spectrometry award
Cathy Costello was recently awarded the 2017 Award for a Distinguished Contribution in Mass Spectrometry by the American Society for Mass Spectrometry. This award recognizes a singular significant achievement and was for her pioneering contributions to the development of tandem mass spectrometry of glycans and glycoconjugates. Addtional details of the award can be found on the ASMS web page. Congratulations Cathy!
Cathy Costello Elected as AAAS Fellow
Co
ngratulations to Cathy Costello for her election as an AAAS Fellow. This is a well-deserved recognition of Dr. Costello’s contributions to mass spectrometry, particularly developing and applying new technologies to advance medicine and increase understanding of biology, and for service to the global community.
Please read article in BUSM News and the full story at the AAAS.
Kshitij Khatri presents at Gordon Research Conference in Italy
Congratulations to Kshitij Khatri (KK) in the Zaia lab for recently presenting at the Glycobiology Gordon Research Conference and Seminar titled "Glycans as Mediators of Interactions Between Molecules, Cells and Organisms". The glycobiology GRC is a unique setting that brings together experts from all fields relevant to carbohydrates in biological systems including synthetic chemistry, structural and systems biology, glycoanalytics and glycoinformatics. KK gave a talk on his research entitled: "Glycoproteomics of Influenza A Virus Hemagglutinin: Correlating Glycan Macro- and Micro-Heterogeneity with Virus Evolution and Interactions with the Host Immune System" and also served as a discussion leader for session, "Chemical and Molecular Analysis of Glycosylation in Health and Disease".
Dr. Catherine Costello named 2011 American Chemical Society Fellow
The American Chemical Society (ACS) inducted BUSM Professor of Biochemistry and Director of the Mass Spectrometry Resource Catherine E. Costello, PhD as an ACS Fellow during the Society’s recent national meeting in Denver. Dr. Costello joins more than 200 distinguished scientists in receiving this honor. Selection is based on outstanding accomplishments in chemistry and important contributions to ACS, the world’s largest scientific society.
Complete details here:
Dr. Costello Receives Outstanding Achievement Award in Mass Spectrometry
Congratulations to Dr. Cathy Costello for receiving the 2010 Frank H. Field and Joe L. Franklin Award for Outstanding Achievement in Mass Spectrometry from the American Chemical Society. This is a tremendous honor and a recognition Cathy’s pioneering contributions to the mass spectrometry of biomolecules, especially carbohydrates and glycoproteins. A link to the B.U. news announcement is below.