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Carboxypeptidase Y Assisted Disulfide-Bond Identification with Linearized Database Search
Jiali Qiang1,2, Zhimin Xu1,2, Yunxia Li1
1Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 100 Haike Road, Pudong, Shanghai 201210, China.
Analytical Chemistry
|November 4, 2021
Summary
We developed carboxypeptidase Y assisted disulfide-bond identification (CADI) to enrich disulfide-linked peptides for mass spectrometry analysis. CADI simplifies complex samples, improving disulfide bond identification in proteins.
Area of Science:
- Biochemistry
- Proteomics
- Analytical Chemistry
Background:
- Disulfide bonds are crucial post-translational modifications regulating protein structure and stability.
- Analyzing disulfide bonds via mass spectrometry is challenging due to difficulties in separating linked peptides and interpreting complex data.
Purpose of the Study:
- To develop a novel strategy for efficient disulfide bond identification.
- To reduce sample complexity and facilitate analysis using standard protein database search engines.
Main Methods:
- Developed carboxypeptidase Y assisted disulfide-bond identification (CADI) strategy.
- CADI depletes linear peptides, enriching disulfide-bonded peptides.
- Data analyzed using standard search engines like Mascot and MaxQuant.
Main Results:
- CADI significantly reduces sample complexity by depleting ~90% of linear peptides.
- The method sensitively identifies disulfide bonds in peptides and proteins.
- Achieved improvement in disulfide bond analysis, though in-depth coverage on complex lysates is still limited.
Conclusions:
- CADI is an effective method for enriching and analyzing disulfide-linked peptides.
- The strategy simplifies mass spectrometry data interpretation for disulfide bond analysis.
- CADI shows potential for large-scale proteomic analysis of disulfide bonds and other cross-linked peptides.
