Remodeling of glycans using glycosyltransferase genes.
نویسندگان
چکیده
Remodeling of glycans on the cell surface is an essential technique to analyze cellular function of lectin-glycan ligand interaction. Here we describe the methods to identify the responsible enzyme (glycosyltransferase) regulating the expression of the glycan of interest and to modulate the glycan expression by overexpressing the glycosyltransferase gene. For the identification of the responsible enzyme, we introduce a new method, CIRES (correlation index-based responsible-enzyme gene screening), that consists of statistical comparison of glycan expression profile obtained by flow cytometry and gene expression profile obtained by DNA microarray.
منابع مشابه
Prediction of Glycan Structures from Glycosyltransferase Expression Profiles
Glycans, which attach to some lipids and Asn/Ser/Thr residues of proteins, draw attentions as the third biological macromolecule next to DNA and protein, since glycans play key roles in the embryogenesis, immunity and diseases. Glycans consist of carbohydrate sugars and their derivatives such as glucose (Glc), mannose (Man), N-acetyl-glucosamine (GlcNAc) and N-acetyl neuraminic acid (Neu5Ac), a...
متن کاملA Gene of the β3-Glycosyltransferase Family Encodes N-Acetylglucosaminyltransferase II Function in Trypanosoma brucei*
The bloodstream form of the human pathogen Trypanosoma brucei expresses oligomannose, paucimannose, and complex N-linked glycans, including some exceptionally large poly-N-acetyllactosamine-containing structures. Despite the presence of complex N-glycans in this organism, no homologues of the canonical N-acetylglucosaminyltransferase I or II genes can be found in the T. brucei genome. These gen...
متن کاملIdentification and Functional Characterization of a Highly Divergent N-Acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei
Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. However, we previously reported a family of puta...
متن کاملCHO cells provide access to novel N-glycans and developmentally regulated glycosyltransferases.
Chinese hamster ovary (CHO) cells express only a subset of the glycosyltransferases activities known to exist. They do not express several fucosyltransferases, galactosyltransferases, sialyltransferases or N-acetylglucosaminyltransferases. However, following mutagenesis or transfection with large amounts of DNA, rare mutants that express a transferase activity de novo have been obtained. The fi...
متن کاملActivation of murine CD4+ and CD8+ T lymphocytes leads to dramatic remodeling of N-linked glycans.
Differentiation and activation of lymphocytes are documented to result in changes in glycosylation associated with biologically important consequences. In this report, we have systematically examined global changes in N-linked glycosylation following activation of murine CD4 T cells, CD8 T cells, and B cells by MALDI-TOF mass spectrometry profiling, and investigated the molecular basis for thos...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Methods in molecular biology
دوره 1200 شماره
صفحات -
تاریخ انتشار 2014