Lysosomal lumen and membrane proteins, as well as non-lysosomal proteins involved in lysosomal function, vary in response to different physiological and pathological conditions. Our engineers have developed several advanced techniques for global analysis of lysosomal proteome dynamics. Here, CD BioSciences provides an effective service to analyze changes in the lysosomal proteome in lysosomal dysfunction diseases.
Background of Lysosomal Lipidome
Since many human diseases are caused by alterations in lysosomes and lysosomal proteins, the lysosomal system is of great importance in biomedicine as alterations in lysosomes. Protein expression analysis is central to the understanding of lysosomal physiology and regulation. Recent advances in the field of proteomics have provided powerful experimental tools for global analysis of lysosomal protein expression. In addition, tandem mass spectrometry-based (LC-MS/MS) can easily capture information on lysosomal composition, expression levels, and modifications. In lysosomal disorders, lysosomal proteins are in a constant state of change and the kinetics of synthesis and degradation are closely related to the function and regulation of lysosomal proteins. Therefore, there is a strong interest in developing proteomic approaches that can analyze the dynamics of lysosomal protein turnover on a global scale.
Fig. 1. How gene mutations in lysosomal enzymes and non-enzymatic lysosomal proteins could lead to LSDs. (Platt F M, et al., 2012)
Our Lysosomal Proteome Changes in Lysosomal Dysfunction Diseases Analysis Services
Lysosomal proteomics has emerged as an important tool for understanding lysosomal dysfunction and LSD pathophysiology in human disease. Based on our ability to rapidly extract high-purity lysosomes, CD BioSciences focuses on lysosomal protein changes from different pathological conditions and cellular states through lysosomal proteome analysis.
Here, we used a proteomic approach to analyze lysosomal proteome changes in animal models of various diseases, including Niemann-Pick disease type C (NPC), Batten's disease, a-mannosidosis, Krabbe disease, Mucolipid Deposition III, Ceroid Lipofuscinosis, etc. With this strategy, we have successfully identified more than 200 proteins, including bona fide lysosomes and lysosome-associated proteins.
- Changes of the lysosomal proteome in NPC
NPC leads to lysosomal cholesterol accumulation. We analyzed changes in the lysosomal proteome of brain tissue by LC-MS/MS, and by knocking out cholesterol transporter protein 1 or 2 (NPC1/NPC2) in nasopharyngeal carcinoma cells of mice. - Changes of the lysosomal proteome in Batten disease
Batten disease is caused by mutations in Battenin (CLN3). We analyzed the changes of the lysosomal proteome by combining SILAC, SPIONs and LC-MS/MS techniques to knock out CLN3. - Changes of the lysosomal proteome in a-mannosidosis
a-mannosidosis is caused by a defect in lysosomal a-D-mannosidase. We used MALDI-MS, 2D-PAGE for protein identification of trimers in mouse liver to reflect changes in the lysosomal proteome in mice with a-mannosidosis.
Our Advantages
- Combination of multiple advanced techniques, such as 2D-PAGE, MALDI-MS, SILAC, SPIONs, LC-MS/MS, gene knockout, etc.
- Experienced team.
- A potentially powerful method for global analysis of lysosomal proteome dynamics.
- Multiple time-point kinetics of lysosomal protein turnover can be analyzed in a single LC-MS/MS run.
- Dramatically reduce the cost and complexity of dynamic proteomics experiments.
- Complete detailed experimental evidences.
We look forward to collaborating with you. You are always welcome to engage in discussions with us at any point of the project. If you are interested in our services, please feel free to contact us for more information.
Reference
- Platt F M, Boland B, van der Spoel A C. (2012) Lysosomal storage disorders: The cellular impact of lysosomal dysfunction[J]. Journal of Cell Biology. 199(5): 723-734.