Mass Spectrometry Determination of Amino Acid Sequences
Mass spectrometry has become the primary method for determining amino acid sequences, particularly peptide and protein sequences. By analyzing the mass spectrometry fragments of the generated peptides, the amino acid sequence of the original peptide or protein can be inferred.
1. Basic Process and Principles of Mass Spectrometry for Determining Amino Acid Sequences
1. Sample Preparation:
Protein Digestion: Large proteins are first cleaved into smaller peptides using specific proteases, such as trypsin.
Peptide Purification: Peptides are purified using HPLC or other chromatographic methods.
2. Peptide Ionization:
In the mass spectrometer, peptides are first ionized. Common ionization methods include electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI).
3. Tandem Mass Spectrometry (MS/MS) Analysis:
The selected precursor ions enter a collision chamber and collide with gas, generating a series of fragment ions. By analyzing these fragment ions, information about the amino acid sequence of the original peptide can be obtained. For example, b-ion series and y-ion series represent the N-terminal and C-terminal sequences of the peptide.
4. Data Analysis:
The collected mass spectrometry data needs to be processed and interpreted using specialized software. The software usually matches the data with known protein databases to infer possible amino acid sequences.
5. Considerations:
The accuracy of determining amino acid sequences is affected by various factors, including the quality of the sample, the performance and resolution of the mass spectrometer, and the methods of data processing and interpretation.
BiotechPack, A Biopharmaceutical Characterization and Multi-Omics Mass Spectrometry (MS) Services Provider
Related Services:
Protein N/C-Terminal Sequencing
N/C-Terminal Sequence Analysis
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