AQUA Ribosomal Proteomic Analysis Service
Are you grappling with the complexities of understanding protein synthesis dynamics, identifying specific ribosomal proteins (RPs) alterations in disease, or seeking accurate quantification of ribosomal components? Our absolute quantification (AQUA) ribosomal proteomic analysis service provides a robust solution for researchers and pharmaceutical companies aiming to meticulously characterize the ribosomal proteome. By delivering quantitative insights into these essential cellular machinery, Creative Biolabs equips your research endeavors with unparalleled precision to identify novel therapeutic targets, validate disease biomarkers, and unravel intricate biological pathways.
Background
The ribosome plays a central role in gene expression regulation. While traditionally viewed as static entities, ribosomes exhibit remarkable heterogeneity in their protein composition and post-translational modifications (PTMs), influencing their translational fidelity and specificity. AQUA proteomics, leveraging stable isotope-labeled synthetic peptides and mass spectrometry (MS), offers an indispensable tool for precisely quantifying these ribosomal components. This targeted approach provides crucial insights into how changes in ribosomal proteome composition contribute to diverse cellular processes and disease states, forming the bedrock for advanced translational research and drug discovery.
Fig.1 Workflow in a bottom-up proteomics experiment.1
Pivotal Process
-
Sample Preparation and Ribosome Isolation
Client-provided samples undergo rigorous preparation, including lysis and differential centrifugation or sucrose cushion isolation to obtain highly purified ribosomal fractions. -
Protein Digestion and Internal Standard Spiking
RPs are denatured and then undergo enzymatic digestion to produce peptides. Crucially, our precisely quantified, stable isotope-labeled synthetic peptides corresponding to your target RPs are spiked into the sample at known concentrations. These internal standards are designed to mimic the native peptides, yet they can be distinguished through MS.
-
LC-MS/MS Analysis (SRM/PRM)
The peptide mixture is separated by high-resolution liquid chromatography (LC) and then introduced into a state-of-the-art tandem mass spectrometer. Using selected reaction monitoring (SRM) or parallel reaction monitoring (PRM), our instruments are specifically tuned to detect and quantify the native and AQUA peptides, leveraging their identical chromatographic retention times and fragmentation patterns, differentiated only by their mass-to-charge ratio.
-
Data Analysis and Absolute Quantification
Sophisticated bioinformatics tools are employed to process the raw MS data. By comparing the signal intensity of the native ribosomal peptides to their corresponding spiked-in AQUA internal standards, the absolute molar concentration of each target RP in your original sample is accurately calculated.
-
Comprehensive Report GenerationThe final stage involves compiling all results into a detailed, easy-to-understand report.
Core Highlights
- Over Two Decades of Expertise
With over twenty years in protein analysis and ribosomal biology, our team has amassed extensive knowledge and practical skills. We've tackled numerous challenges, enhancing problem-solving abilities. Working with us means leveraging this expertise for more successful research.
- Cutting-Edge MS Platforms
We use top-notch high-resolution LC-MS/MS systems. These offer unmatched sensitivity to detect faint traces and high specificity to avoid misidentifications, providing precise data for reliable conclusions.
- Exceptional Reproducibility
Reproducibility is key in our work. Our methods are carefully designed with strict quality control at each step, ensuring consistent and replicable quantitative data that adds credibility to your findings.
- Precise PTM Characterization
Quantifying PTMs on RPs is tough but vital. We've developed special techniques to accurately identify and measure PTMs, offering insights into protein regulation and potential therapeutic targets.
Applications
- Understanding Translational Control Mechanisms
Precisely quantify changes in RP levels and modifications under different cellular conditions, such as stress, growth, and differentiation, to elucidate mechanisms regulating protein synthesis efficiency and specificity.
- Drug Target Validation
Characterize the effects of drug candidates on RP composition and PTMs, pinpointing specific RPs or their modifications as potential therapeutic targets, and assessing off-target effects.
- Ribosomopathy Research
Quantify specific RP deficiencies or alterations linked to ribosomopathies, aiding in diagnosis, prognosis, and the development of targeted therapies.
- Investigating Pathogen-Host Interactions
Analyze changes in host ribosomal proteome upon infection, or characterize pathogen-encoded ribosomal-like proteins, providing insights into viral or bacterial manipulation of host translational machinery.
- Cellular Quality Control and Aging Studies
Monitor the integrity and composition of ribosomes during cellular aging or in response to cellular stressors, contributing to a deeper understanding of proteostasis and age-related diseases.
FAQs
Q: How does AQUA ribosomal proteomic analysis compare to traditional WB for RP quantification?
A: While WB offers relative quantification, our AQUA service provides absolute quantification of RPs. This means you receive precise molar concentrations, enabling more accurate comparisons across samples and experiments. AQUA also offers higher multiplexing capabilities and superior sensitivity for low-abundance proteins, particularly valuable when analyzing complex ribosomal PTMs.
Q: Can your service differentiate between different RP isoforms or their PTMs?
A: Absolutely. Our AQUA peptides are specifically engineered to target defined protein sequences, including those harboring PTMs, such as phosphorylation, acetylation, and methylation. This allows for the precise quantification of distinct RP isoforms and their modified states, providing a granular understanding of ribosomal heterogeneity and function.
Q: Is the AQUA ribosomal proteomic analysis service cost-effective for large-scale studies?
A: While the initial setup for AQUA can be more involved than some other methods, its unparalleled accuracy, sensitivity, and quantitative precision often lead to significant long-term cost savings by reducing the need for repeat experiments and providing more definitive biological insights. We offer competitive pricing and flexible project scopes to accommodate various research budgets, and we're happy to provide a detailed quote based on your specific requirements.
Our AQUA ribosomal proteomic analysis service offers a high-precision quantitative platform for systematic dissection of ribosomal biology complexity, enabling unparalleled insights into translational regulation mechanisms. By leveraging advanced MS and stable isotope-labeled standards, we empower researchers to gain unprecedented insights into translational control, ribosome heterogeneity, and the role of RPs in health and disease. Our commitment to accuracy, scientific excellence, and personalized service makes us your ideal partner for accelerating discovery in this critical area of biochemistry. Contact us now!
Reference
- Dupree, Emmalyn J., et al. "A critical review of bottom-up proteomics: the good, the bad, and the future of this field." Proteomes 8.3 (2020): 14. DOI: 10.3390/proteomes8030014. Distributed under Open Access license CC BY 4.0, without modification.
(USA)
(UK)
(Germany)
