What is Amino Acid Composition?
Amino acids are the fundamental building blocks of proteins, and they play crucial roles in processes such as neurotransmitter transport and biosynthesis. Currently, 22 amino acids have been identified as constituents of proteins. The amino acid composition of a protein refers to the percentage each amino acid occupies among all the amino acids comprising that protein. Sometimes referred to as molar percentage, it is calculated by dividing the number of each standard amino acid by the total number of amino acids in the protein chain or molecule.
Amino acid composition analysis is the gold standard for accurately quantifying samples such as peptides, proteins, antibodies, etc. Typically, it is desirable to determine the amino acid composition of a sample before obtaining its sequence information. This aids in identifying errors in the sequencing process and allows for necessary corrections to be made. Understanding the amino acid composition of a sample also helps in determining which proteinase is suitable for the sample's hydrolysis process. Additionally, it assists in identifying the presence of low-level non-standard amino acids (e.g., norleucine) in proteins.
Creative Proteomics offers amino acid analysis services for samples including amino acids, peptides, polypeptides, proteins, and antibodies. Our services involve both qualitative and quantitative determination of the amino acid composition in the samples.
Amino Acid Composition Analysis in Creative Proteomics
Amino Acid Analysis Process:
1. Sample Preparation: We begin by carefully preparing your protein sample to ensure its purity and integrity, a critical step in obtaining accurate results.
2. Hydrolysis: The protein sample undergoes hydrolysis, breaking down the peptide bonds to release individual amino acids.
3. Derivatization (if necessary): Depending on the nature of the sample and the analysis requirements, derivatization may be employed to enhance the detection sensitivity of specific amino acids.
4. Analysis by HPLC or Mass Spectrometry: The hydrolyzed and, if applicable, derivatized amino acids are then separated and quantified using high-performance liquid chromatography (HPLC) or mass spectrometry.
5. Data Analysis and Reporting: Our bioinformatics team analyzes the data and generates a comprehensive report, presenting the amino acid composition of your protein in a clear and interpretable format.
Analytical Technology Platform:
Mass spectrometry platforms:
- MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry): Utilizing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the analysis and identification of amino acids and small peptides.
- ESI-MS (Electrospray Ionization Mass Spectrometry): Electrospray ionization, often coupled with liquid chromatography (LC-MS), is employed for the analysis and quantification of the amino acid composition of proteins and peptides.
Chromatography platforms:
- Ion Exchange Chromatography: Utilizing the charge characteristics of amino acids for separation.
- RP-HPLC (Reverse Phase High-Performance Liquid Chromatography): Typically combined with mass spectrometry for accurate amino acid quantification analysis.
Automatic amino acid analyzer
Advantages of Amino Acid Composition Analysis
Profound Expertise in Protein Analysis: Benefit from our extensive knowledge and experience in the realm of protein analysis, ensuring a thorough examination of intricate compositions.
High Sensitivity and Accuracy for Diverse Protein Modifications: Our service excels in pinpointing proteins with a spectrum of post-translational modifications, providing the high sensitivity and accuracy essential for comprehensive insights.
Rapid Turnaround Time: Experience efficiency with a swift turnaround time of 5-7 days, ensuring a comprehensive report is delivered promptly to meet your timelines.
Customized Service: Tailored to your unique requirements, our service offers customization based on your sample type and size. Optimized protocols are meticulously deployed, ensuring accuracy and relevance in every analysis.
Samples for Amino Acid Composition Analysis
Sample Format: Samples can be in the form of solids or solutions.
Sample Amount:
- For dry protein/peptide samples: A minimum of 100 µg or more, with a concentration ranging approximately from 0.5 to 2 mg/mL.
- For liquid materials: A volume exceeding 100 μL, with a minimum concentration of 0.5 mg/mL.
Feel free to reach out to us should you have any additional requirements for your samples.
Case Extraction and Characterization of Type I Collagen from Pearl Mullet (Alburnus tarichi Güldenstädt, 1814) Skin and Bone
Background
Pearl mullet (Alburnus tarichi) is a cyprinid fish species endemic to Turkey's eastern region, particularly Lake Van. Due to its unique habitat and significant decline in population, the extraction and characterization of collagen from pearl mullet skin and bone become crucial for potential applications in various industries.
Samples
The study utilized pearl mullet specimens, focusing on both skin (ASC-S) and bone (ASC-B) components. The samples were obtained from the local fish market, with subsequent cleaning, separation of skin and bones, and freezing for preservation.
Technical Methods
Sample Preparation: The collagen samples were prepared following a modified method based on Nagai and Suzuki (2000a). Procedures were conducted at temperatures not exceeding +4 °C.
Characterization Techniques:
- Collagen Yields Calculation: The collagen yields were calculated based on the dry weight of the initially used material.
- Differential Scanning Calorimetry (DSC): Analysis of thermal stability, including Tmax and enthalpy values.
- X-Ray Diffraction (XRD): Determination of crystal structures.
- Fourier Transform Infrared Spectroscopy (FTIR): Analysis of collagen spectra.
- UV-Vis Measurement: Assessment of collagen purity through UV spectra.
- Scanning Electron Microscopy (SEM): Visualization of morphological structures.
- Amino Acid Composition: Hydrolysis under vacuum, followed by HPLC analysis for amino acid composition.
Statistical Analysis: One Way ANOVA used to determine differences between groups.
Results
Collagen Yield: ASC-S exhibited a higher yield than ASC-B, with variations observed compared to other fish species in the literature.
Thermal Stability (DSC): Lower imino acid content in pearl mullet contributed to better thermal stability, aligning with subtropical and tropical fish collagen characteristics.
XRD Analysis: Preservation of triple helix structure confirmed in both ASC-S and ASC-B, consistent with findings in various fish species.
FTIR Spectra: Shifts in Amide A and other bands indicated the presence of hydrogen-bonded hydroxyl groups, confirming type I collagen.
UV-Vis Spectroscopy: Distinctive absorption peaks at 230-232 nm confirmed collagen's type I nature, with absence of tryptophan contributing to observed absorbance range.
SEM Images: Morphological characteristics revealed porous structures with dense, irregular surfaces, supporting the classification of type I collagen.
Reference
- GÖÇER, Mustafa, Yasemen YANAR, and Muhsin AYDIN. "A study on amino acid composition and distinctive traits of collagen isolated from the skin and bone of an endemic pearl mullet species, Alburnus tarichi (Güldenstädt, 1814)." GSC Biological and Pharmaceutical Sciences 25.2 (2023): 353-364.
