An Efficient Method for Protein Sequence and PTM Characterization with Complete Coverage

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	An Efficient Method for Protein Sequence and PTM Characterization with Complete Coverage
	Ang, I. L.; Wei, R.; Lam, M. M. T.; Lei, K.; Zhang, P.; Poon, C. W.
	2018-06-08
Source Publication	5th Macau Symposium on Biomedical Sciences 2018
Abstract	n the field of proteomics, protein identification and characterization of post-translational modifications (PTMs) require proteolysis of proteins into peptides for fragmentation by tandem mass spectrometry followed by computational mapping of peptide sequences. Trypsin is the most commonly used protease though it provides limited protein sequence coverage owing to its substrate specificity to lysine and arginine. The limited protein sequence coverage issue can be solved by undertaking an approach involving multiple independent enzymatic digestions with two or more digestive enzymes that cleave at different residues to generate peptides which contains overlapping amino acid sequences across the protein sequence. Unfortunately, such approach is tedious, time consuming, and substantially increases the amount of protein required for analysis of protein sequence and PTMs. In the present study, we developed a new method to obtain enhanced protein sequence coverage through generation of multiple overlapping peptides. The method was shown to be able to achieve 100% protein sequence coverage for single protein or protein within a mixture, and was applicable to low amount of proteins (62.5ng) or large proteins (>150kDa). Up to 99.9% protein sequence coverage was obtained when the method was applied to sequence a 6-protein mixture. Moreover, protein PTMs, such as phosphorylation, glycosylation and protein N-terminal modification could be characterized at the same time. Comparing to existing methods, our new method is fast and sensitive with the whole process requiring only 3 hours.
Keyword	Proteomics protein sequence post-translational modification mass spectrometry
Language	英語English
The Source to Article	PB_Publication
PUB ID	41819
Document Type	Conference paper
Collection	DEPARTMENT OF BIOMEDICAL SCIENCES
Corresponding Author	Poon, C. W.
Recommended Citation GB/T 7714	Ang, I. L.,Wei, R.,Lam, M. M. T.,et al. An Efficient Method for Protein Sequence and PTM Characterization with Complete Coverage[C], 2018.
APA	Ang, I. L.., Wei, R.., Lam, M. M. T.., Lei, K.., Zhang, P.., & Poon, C. W. (2018). An Efficient Method for Protein Sequence and PTM Characterization with Complete Coverage. 5th Macau Symposium on Biomedical Sciences 2018.

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