Integrated Technology Resource for Biomedical Glycomics

Integrated Technology Resource for Biomedical Glycomics

Research Emphasis

The primary focus of the Integrated Technology Resource for Biomedical Glycomics is the development of technologies to analyze and influence glycoprotein and glycolipid glycosylation found in animal cells. The resource has particular interest in developing tools and techniques to analyze glycosylation during animal cell differentiation and development as well as during oncogenesis and tumor progression. The broad goal of these studies is to develop the means to describe the patterns of glycosylation changes during these differentiation events and determine whether glycosylation inhibitors can influence differentiation pathways.

Current Research

The resource focuses on the early stages of embryonic mouse stem cell differentiation as the primary system of study for technology development. Glycoprotein and glycolipid glycosylation is being characterized during differentiation of these stem cells into cells in the ectodermal primitive layer and during further differentiation into neuronal precursor cells. Glycoprotein analysis will identify N- and O-linked glycans, and the proteins to which these glycans are attached, using liquid chromatography/mas spectrometry (MS)-MS proteomic techniques coupled to serial lectin affinity chromatography schemes. In addition, sphingolipid and glycosphingolipid structures will be characterized to identify changes during differentiation. Changes in particular glycosyltransferases during differentiation will be determined by glycochip analysis, followed by quantification by kinetic polymerase chain reaction experiments. A bioinformatics component will integrate the glycosylation information obtained and will synergize with other glycodatabases being developed around the world.

Resource Capabilities

Instruments

Hewlett-Packard gas chromatography-MS instruments, DIONEX HPAEC system, Applied Biosystems Voyager DE matrix-assisted laser desorption ionization (MALDI)-MS mass spectrometer, Metrohm-Peak HPAEC system, and Thermo Finnegan LCQ and LTQ Advantage mass spectrometers.

Software

Innovative software is currently under development.

Training Opportunities and Workshops

Services offered include initial characterization of glycoprotein and glycosphingolipid glycans, including estimation of sample purity, glycosyl composition analysis, glycosyl linkage analysis, and molecular weight determinations using gas-liquid chromatography, electrospray ionization, fast-atom bombardment, MALDI, time-of-flight, and tandem MS. Proteomic analysis of N-glycosylated glycopeptides also is available. More sophisticated, customized, or nonroutine services can be individually designed and performed as a collaborative investigation. Several workshops emphasizing methodology for glycoconjugate analysis are offered each year, and participants come from both industry and academia.