A validated collection of mouse monoclonal antibodies to human glycosyltransferases functioning in mucin-type O-glycosylation
| Autor: | Shengjun Wang, Zhang Yang, Yoshiki Narimatsu, Katrine T. Schjoldager, Christoffer K. Goth, Sarah L. King, Tongzhong Ju, Lars Hansen, Eric P. Bennett, María F Festari, Malene Bech Vester-Christensen, Ulla Mandel, Catharina Steentoft, Kelley W. Moremen, Ida Signe Bohse Larsen |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Glycosylation
medicine.drug_class Monoclonal antibody Biochemistry 03 medical and health sciences chemistry.chemical_compound symbols.namesake Mice Antibody Specificity medicine Animals Humans Secretory pathway 030304 developmental biology 0303 health sciences Chemistry 030302 biochemistry & molecular biology Mucins Antibodies Monoclonal Glycosyltransferases Reproducibility of Results Golgi apparatus Subcellular localization Zinc finger nuclease Cell biology Secretory protein HEK293 Cells Proteome symbols Original Article |
| Zdroj: | Glycobiology |
| ISSN: | 1460-2423 |
| Popis: | Complex carbohydrates serve a wide range of biological functions in cells and tissues, and their biosynthesis involves more than 200 distinct glycosyltransferases (GTfs) in human cells. The kinetic properties, cellular expression patterns and subcellular topology of the GTfs direct the glycosylation capacity of a cell. Most GTfs are ER or Golgi resident enzymes, and their specific subcellular localization is believed to be distributed in the secretory pathway according to their sequential role in the glycosylation process, although detailed knowledge for individual enzymes is still highly fragmented. Progress in quantitative transcriptome and proteome analyses has greatly advanced our understanding of the cellular expression of this class of enzymes, but availability of appropriate antibodies for in situ monitoring of expression and subcellular topology have generally been limited. We have previously used catalytically active GTfs produced as recombinant truncated secreted proteins in insect cells for generation of mouse monoclonal antibodies (mAbs) to human enzymes primarily involved in mucin-type O-glycosylation. These mAbs can be used to probe subcellular topology of active GTfs in cells and tissues as well as their presence in body fluids. Here, we present several new mAbs to human GTfs and provide a summary of our entire collection of mAbs, available to the community. Moreover, we present validation of specificity for many of our mAbs using human cell lines with CRISPR/Cas9 or zinc finger nuclease (ZFN) knockout and knockin of relevant GTfs. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|