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Complete enzymic digestion of acidic proteins
International Journal of Peptide and Protein Research
|January 1, 1976
Summary
A new enzymatic hydrolysis method effectively breaks down resistant acidic proteins. This approach accurately quantifies amino acids, including those unstable under acid conditions, for comprehensive protein analysis.
Area of Science:
- Biochemistry
- Proteomics
- Analytical Chemistry
Background:
- Acidic proteins often resist complete enzymatic hydrolysis.
- Standard acid hydrolysis methods degrade certain unusual amino acids.
- A universal enzymatic hydrolysis technique is needed for accurate protein analysis.
Purpose of the Study:
- To develop and validate a complete enzymatic hydrolysis method for acidic proteins.
- To ensure accurate recovery of all amino acids, including acid-labile ones.
Main Methods:
- Utilized a sequential enzymatic approach involving subtilisin, leucine amino-peptidase, prolidase, and carboxypeptidase C.
- Applied the method to four distinct acidic proteins.
Main Results:
- Achieved complete hydrolysis of the target acidic proteins.
- Observed excellent agreement between recovered amino acid content and known sequences.
- Demonstrated the method's efficacy for proteins containing unusual amino acids.
Conclusions:
- The described multi-enzyme cocktail provides a robust method for complete protein hydrolysis.
- This technique overcomes limitations of acid hydrolysis and is applicable to diverse protein types.
- Enables accurate amino acid quantification essential for proteomics and protein characterization.