Glycoproteins are delivered to lysosomes for catabolism by extracellular endocytosis or intracellular autophagy. Lysosomal catabolism of glycoproteins is currently part of the normal turnover of cellular components and cellular homeostasis of glycosylation. CD BioSciences' expertise in lysosomes provides strong support for glycosylation analysis of lysosomal glycoproteins. Our advanced mass spectrometry (MS) technology platform enables complete glycopeptide analysis to ensure your 100% satisfaction.
Lysosomal Glycoproteomics
Protein glycosylation, which involves the co- or post-translational addition of glycans to proteins, is a crucial protein modification in health and disease. The vast majority of lysosomal proteins are highly glycosylated. Once inside the lysosome, glycoproteins are broken down by a combination of proteases and glycosidases, characteristic of soluble lysosomal hydrolases. Lysosomal catabolism of glycoproteins has become an important part of cellular glycosylation homeostasis, and many diseases caused by defects in glycoprotein lysosomal catabolism have emerged. Asparagine-linked (N-linked) and serine/threonine-linked (O-linked) glycans have attracted great interest in mammalian lysosomes.
Fig. 1. The role of lysosomal catabolism of glycoproteins in the cellular homeostasis of glycosylation. (Winchester B, 2005)
Glycosylation Analysis Services for Lysosomal Glycoproteins
Characterization of intact glycopeptides is an attractive analytical strategy to study protein site-specific glycosylation. CD BioSciences uses liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based analysis of intact glycopeptides for bottom-up glycoproteomics, allowing comprehensive analysis of lysosomal glycoprotein glycosylation events, including Peptide sequences, glycan structures and modification sites. In addition, we used glycosylases to analyze N- and O-glycans attached to lysosomal proteins.
Here, our engineers can characterize thousands of intact N-glycopeptides and O-glycopeptides in lysosomal glycoproteomics experiments. We have developed a detailed protocol for lysosomal glycoproteomics research, the process is as follows:
- Enrichment of glycopeptides based on chemical and biological affinity.
- The glycopeptide sequence was determined based on MS.
- Depending on the type of glycosylation under consideration, locate the glycan attachment site or sites in the peptide.
- Relative quantification of glycosylation in different samples.
Advantages of Our Services
- Advanced MS technology platforms including matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry, nano- electrospray ionization mass spectrometry/mass spectrometry (nano-ESI-MS-MS).
- Experienced professional team.
- Cutting-edge computational tools for glycopeptide identification.
- Complete data storage.
- Different types of carbohydrate residues or chains are removed with specific glycosidases.
- Helps you determine that the antibody of your choice reacts identically to the glycosylated and deglycosylated forms of the glycoprotein of interest.
Our professional services for N- and O-glycosylation analysis of lysosomal glycoproteins have been well received by customers. Our highly skilled and dedicated scientific staff ensures that the most appropriate method and technology is selected for each specialized lysosomal project. If you have any special requirements about our services, please feel free to contact us. We are looking forward to working together with your attractive projects.
References
- Winchester B. (2005) Lysosomal metabolism of glycoproteins[J]. Glycobiology. 15(6): 1R-15R.
- Tokhtaeva E, Mareninova O A, Gukovskaya A S, et al. (2017) Analysis of N-and O-glycosylation of lysosomal glycoproteins[M]// Lysosomes. Humana Press, New York, NY. 35-42.