Peptidoglycan Structure Analysis
CD BioGlyco has rich experience in peptidoglycan structural analysis. Our multiple technology platforms and well-trained researchers can help customers get results quickly and accurately in the case of low-abundance samples. We have confidence to be your essential research assistant in the field of glycobiology.
Peptidoglycan (PG, also known as murein) exists in the cell walls of gram-positive bacteria and gram-negative bacteria in eubacteria. The backbone of peptidoglycan is a polysaccharide chain formed by two sugar derivatives: N-acetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc) that alternately connected by β-1,4-glycosidic bonds. Two parallel sugar chains are connected laterally to form a network of peptidoglycan. PG is essential for maintaining cell integrity because of its ability to withstand turgor pressure. Any external force that affects its biosynthesis or degradation will interfere with cell viability. Therefore, PG is one of the main targets of antibacterial drugs. The study of peptidoglycan structure also plays a vital role in understanding the immune response, phage susceptibility and serological behavior, as well as the classification and identification of bacteria. Although the structure and architecture of PG have been studied for a long time, its detailed 3D structure is still an open question.
CD BioGlyco provides a one-stop service to assist in studies from PG separation to structural analysis, and each process is strictly controlled by experienced researchers. The experimental operations are built on our multiple technology platforms, including but not limited to:
Fig.1 Key components of peptidoglycan structural analysis.
Our streamlined workflow ensures a comprehensive and efficient process, from initial consultation to final product delivery. Our team of experts works closely with you to tailor each step to your project's specific needs.
We begin by growing the specified bacterial strain under optimized conditions to ensure high biomass. The cells are then harvested, washed, and prepared for the extraction process.
The prepared bacterial cells are subjected to a carefully selected extraction method (e.g., phenol-water, butanol, or a proprietary method) to release the target carbohydrates from the cell membrane. This crude extract then undergoes preliminary purification to remove major contaminants.
The extract is treated with a specific cocktail of enzymes, such as proteinase K, RNase, and DNase I, to digest and remove contaminating proteins and nucleic acids. This is followed by advanced chromatography to achieve the highest possible purity.
The final purified product is subjected to a rigorous suite of analytical tests. We use HPLC and SDS-PAGE to confirm purity and homogeneity. For detailed structural analysis, we perform mass spectrometry to characterize the precise molecular structure of the carbohydrate.
DOI.: 10.1371/journal.ppat.1010241
Journal: PLoS pathogens
Published: 2022
Results: Salmonella typhimurium undergoes significant peptidoglycan remodeling during its non-proliferative intracellular phase in fibroblasts, characterized by atypical crosslinked muropeptides with stem peptides trimmed at L-Ala-D-Glu(γ) or D-Glu(γ)-meso-diaminopimelic acid motifs. These modifications include a ~30% increase in L, D-crosslinks catalyzed by L, D-transpeptidases LdtD and LdtE, reduced glycan chain length, and an unprecedented terminal D-alanine substitution with alaninol (detected via untargeted MS/MS). Crucially, synthetic tetrapeptides bearing alaninol attenuate NF-κB nuclear translocation in macrophages, revealing a novel immune evasion strategy. This peptidoglycan editing allows intracellular Salmonella to persist by interfering with NOD1/NOD2-mediated immune recognition while residing within host cells.
Building upon the expertise in intricate peptidoglycan purification and structural characterization, our analytical capabilities extend to a diverse range of polysaccharides from various natural sources. This specialized knowledge is directly applied in our comprehensive Polysaccharide Analysis Service, which includes, but is not limited to:
CD BioGlyco provides systematic and one-stop services for peptidoglycan structure research and ensure the continuity and transparency of the experimental process. We will continue to improve our standards to meet customers' glycobiology research needs with high quality.
Customers can contact our employees directly and we will respond promptly. If you are interested in our services, please contact us for more detailed information.
Reference
