Other Soluble Proteins Structure Characterization Service | Cryo-EM

With the rapid advancement of structural biology, accurately determining the three-dimensional structures of proteins has become essential for understanding protein functions and developing novel therapeutics. Beyond traditional targets such as membrane proteins, multiprotein complexes, GPCRs, and ion channels, numerous other types of naturally soluble proteins, including kinases, phosphatases, proteases, DNA polymerases, and transcription factors, are increasingly attracting researchers' attention. These proteins typically exist naturally in soluble form within cells, display good stability, and participate in multiple critical biological processes. Nevertheless, determining their three-dimensional structures remains challenging due to their structural diversity and dynamic characteristics.

 

MtoZ Biolabs addresses this challenge effectively and accurately with the Other Soluble Proteins Structure Characterization Service Based on Cryo-EM, supporting scientific research and pharmaceutical development.

 

Technical Principles

Cryogenic Electron Microscopy (Cryo-EM) has rapidly emerged as a high-resolution structural determination method, capable of achieving near-atomic resolution. Unlike traditional crystallography, Cryo-EM bypasses crystallization, making it highly suitable for naturally soluble proteins, which typically possess flexible structures and good solubility.

 

MtoZ Biolabs employs advanced Cryo-EM equipment and high-performance direct electron detectors, combined with efficient image processing algorithms and a highly experienced professional team. We provide high-quality Other Soluble Proteins Structure Characterization Services based on Cryo-EM, capable of accurately characterizing the structures of diverse soluble proteins.

 

Analysis Workflow

MtoZ Biolabs' Other Soluble Proteins Structure Characterization Service follows a standardized and optimized Cryo-EM workflow:

 

1. Sample Preparation and Quality Assessment: After receiving client samples, we conduct initial quality assessments, including concentration, purity, and homogeneity evaluations, ensuring suitability for Cryo-EM analysis.

 

2. Sample Vitrification: Using specialized vitrification equipment, protein samples are rapidly frozen into amorphous ice, preserving their native conformation and state to the greatest extent.

 

3. Cryo-EM Data Acquisition: High-end Cryo-EM systems are used to acquire two-dimensional projection images of proteins, generating high-resolution raw data.

 

4. Data Processing and 3D Reconstruction: Advanced processing software is utilized for image correction, particle picking, 2D classification, and 3D reconstruction, yielding three-dimensional structural models of proteins.

 

5. Structure Model Construction and Optimization: High-precision protein structural models are constructed and optimized using structural refinement software, providing detailed structural information.

 

6. Results Delivery and Reporting: Comprehensive analytical reports, structural model files, and expert structural interpretations are provided.

 

Service Advantages

1. Professional Equipment and Team Support: MtoZ Biolabs boasts a globally recognized Cryo-EM platform and a team with extensive experience in protein structural characterization.

 

2. High Efficiency and Accuracy: Eliminating crystallization reduces failure risk and accelerates structural characterization processes.

 

3. Wide Applicability: Suitable for various naturally soluble proteins, including kinases, phosphatases, transcription factors, and chaperones, meeting diverse research demands.

 

4. Customized Service: Experimental strategies are flexibly adjusted according to client requirements, ensuring optimal structural determination solutions.

 

Applications

MtoZ Biolabs’ Other Soluble Proteins Structure Characterization Service is broadly applicable to the following fields:

 

1. Drug Discovery and Screening: Structural determination of drug target proteins to guide molecular drug design.

 

2. Signal Pathway Research: Detailed understanding of the structural and functional roles of key kinases and phosphatases in signal transduction.

 

3. Fundamental Biological Research: Revealing mechanisms of transcription factors, proteases, and other proteins in vital biological processes.

 

4. Industrial Enzyme Engineering: Providing structural foundations for engineering enzyme proteins to enhance industrial production efficiency.

 

Case Study

1. High-Resolution Cryo-EM Analysis of the Human CDK-Activating Kinase

This study employs high-resolution cryo-electron microscopy (cryo-EM) to determine the structure of the human CDK-activating kinase (CAK), a critical regulator of cell growth and division. Structures of CAK in its free, nucleotide-bound, and inhibitor-bound states were resolved, reaching resolutions up to 1.8 Å. Detailed structural analyses reveal inhibitor interactions and networks of water molecules within the active site, elucidating mechanisms governing inhibitor selectivity and laying the structural foundation for rational next-generation drug design. The Other Soluble Proteins Structure Characterization Service utilizes high-resolution cryo-EM technology to accurately determine structures of naturally soluble proteins such as kinases, phosphatases, and transcription factors. It effectively captures protein dynamic conformations and ligand-binding interactions, facilitating drug discovery and functional protein studies.

 



Cushing, VI. et al. Nat Commun. 2024.

 Figure 1. High-Resolution Structures of the Human CDK-Activating Kinase

 

2. Cryo-EM Structure of Phospholipase Ce Defines N-terminal Domains and Their Roles in Activity

This study presents a 3.9 Å resolution cryo-EM structure of phospholipase C epsilon (PLCε) in complex with an antigen-binding fragment (Fab), defining critical N-terminal domains. The structure uncovers a pleckstrin homology (PH) domain and four tandem EF-hand domains with specific insertions and interactions with the catalytic core. Structural modeling and functional assays suggest these domains form a continuous membrane-binding surface crucial for basal and G protein-stimulated PLCε activity, providing insights into its autoinhibited and activated states. Other Soluble Proteins Structure Characterization Service leverages cryo-EM technology to elucidate structures of soluble proteins with complex domain organizations, such as enzymes with regulatory regions, capturing conformational dynamics and intramolecular interactions to support functional characterization and structural studies.

 



Samassekou, K. et al. bioRxiv. 2024.

Figure 2. Cryo-EM Reconstruction of the Fab2–PLCe PH-C Complex

 

FAQ

Q1: What requirements should the submitted protein samples meet?

A1: Protein samples should have high purity (recommended >90%), concentrations typically ranging from 1-5 mg/mL, and solutions free from components interfering with Cryo-EM analysis.

 

Q2: How long does the Cryo-EM analysis usually take?

A2: Depending on project complexity and protein properties, the typical project duration is 4-8 weeks. Timelines can also be flexibly adjusted according to urgent client needs.

 

Q3: If sample quality is insufficient, what support does MtoZ Biolabs provide?

A3: We provide preliminary quality assessment reports and optimization recommendations, assisting clients in improving sample preparation procedures.

 

Q4: What forms of data are delivered from Cryo-EM analysis?

A4: We deliver protein 3D structure model files (PDB format), high-resolution structural images, detailed structural analysis reports, and raw data.

 

Q5: Is technical support for structural analysis provided after results delivery?

A5: Yes, our team offers ongoing technical support, including structural interpretation, model optimization suggestions, and guidance for subsequent experimental design.

 

Through MtoZ Biolabs' Other Soluble Proteins Structure Characterization Service, researchers can gain deeper insights into soluble protein structures and functions, driving breakthroughs in pharmaceutical development and life science research.