Protein N-terminal analysis using fast atom bombardment mass spectrometry

Area of Science:

• Analytical Chemistry
• Biochemistry
• Proteomics

Background:

• N-terminal protein sequencing is crucial for protein identification and characterization.
• Dansyl chloride derivatization is a common method for amino acid analysis.
• Limitations exist in the sensitivity and quantitation of traditional methods.

Purpose of the Study:

• To establish Fast Atom Bombardment (FAB) mass spectrometry for identifying and quantifying dansyl amino acids.
• To assess the quantitative capability of FAB mass spectrometry in N-terminal protein analysis.
• To validate the technique using various protein samples.

Main Methods:

• Fast atom bombardment (FAB) mass spectrometry was utilized.
• Dansyl amino acids were prepared from N-terminal protein analysis.
• Dansyl aminobutyric acid served as an internal standard for quantitation.
• Analysis was performed on proteins including bovine serum albumin and ribonuclease A.

Main Results:

• FAB mass spectra of dansyl amino acids exhibited characteristic quasimolecular and fragment ions.
• Quantitative determination of dansyl amino acids was achieved at levels as low as 0.1 nmol.
• The method demonstrated successful N-terminal residue analysis across multiple protein types.

Conclusions:

• FAB mass spectrometry is a sensitive and quantitative technique for dansyl amino acid analysis.
• This method significantly advances N-terminal protein sequencing capabilities.
• The validated approach provides reliable data for protein characterization.