Background of Lysosomal Protein Enrichment
Lysosomes have a decisive role in a wide range of diseases that are often caused by genetic defects in lysosomal proteins or proteins involved in lysosomal biogenesis. Furthermore, the degradation function of lysosomes is carried out within the lysosomal lumen through the synergistic action of more than 60 hydrolases and auxiliary proteins. Over the past decade, scientists have used large-scale mass spectrometry-based methods to study the composition of lysosomal proteins. A variety of comprehensive subcellular isolation methods have been used to purify intact lysosomes, and by characterising the proteomic profile of lysosomes can provide valuable information about their function in normal and disease states. However, density centrifugation has limited resolution for subcellular isolation due to the overlapping physical properties of organelles, often allowing enrichment of organelles but not purification to homogeneity. Therefore, a more precise study of lysosomal proteomics by enrichment of these proteins is essential.
Fig. 1. Strategies for the enrichment of lysosomes and lysosomal membrane proteins. (Muthukottiappan P, et al., 2021)
Our Lysosomal Protein Enrichment Services
Proteins involved in lysosomal function can be classified as soluble proteins in the lumen of the lysosome, membrane proteins, and proteins present in the cytoplasm associated with the lysosomal surface. In addition to enrichment of intact organelles, lysosomal luminal proteins can also be enriched by affinity purification. For many years, CD BioSciences has been working on enriching lysosomal proteins by affinity purification, which helps to improve the accuracy of analytical or preparative subcellular isolation methods for organelle proteomics.
We have successfully identified enriched lysosomal proteins from the lysosomes of rat liver. Here, we used the following affinity chromatography purification strategies for proteomic characterization. We will combine the advantages and disadvantages of each strategy to develop the most effective and cost-effective solution for you.
- Purification of Lysosomal Proteins by Mannose 6-Phosphate
Our affinity purification based on mannose 6-phosphate (Man6-P) modification for greater enrichment of proteins located in the lysosomal compartment can help you identify proteins containing Man6-P. We have now identified 60 known lysosomal proteins, representing almost all of the known proteins containing Man-6-P.
- This method can be applied to almost any type of sample.
- High specificity for lysosomal proteins.
- Limited to a small subset of lysosomal hydrolases.
- Proximity Biotinylation of Lysosomal Membrane and Associated Proteins
We used in vivo proximity biotinylation methods to analyze lysosomal membrane protein interactions and organelle proteomes.
- Labeling can be performed in live cells and animals in a short period of time.
- Allows the study of transient in vivo interactions, which may be lost during enrichment.
- Limited to the outer surface of the lysosomal membrane.
CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.
Reference
- Muthukottiappan P, Winter D. (2021) A proteomic view on lysosomes[J]. Molecular Omics.