Applicability of the Alkylation Chemistry for Chemical C-Terminal Protein Sequence Analysis

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• 作者：Bart Samyn ; Klaas Hardeman ; Johan Van der Eycken ; and Jozef Van Beeumen
• 刊名：Analytical Chemistry
• 出版年：2000
• 出版时间：April 1, 2000
• 年：2000
• 卷：72
• 期：7
• 页码：1389 - 1399
• 全文大小：432K
• 年卷期：v.72,no.7(April 1, 2000)
• ISSN：1520-6882

文摘

We have evaluated the alkylation chemistry first describedsome years ago by Boyd et al. which is now routinelyapplied in a commercial instrument. We have found thatthe low repetitive yields observed during these analysesare due to the formation of a major side product whenalkylating the C-terminal thiohydantoin. This side product,resistant to the chemical cleavage methods currently used,was characterized by NMR experiments in solution. Wefurther demonstrate that chemical C-terminal sequenceanalysis of proteins using the alkylation chemistry isfeasable with low picomole amounts of material. High-sensitivity C-terminal sequencing allows a complementaryapproach by which a protein is first subjected to N-terminal Edman degradation followed by C-terminal sequence analysis, limiting the amount of material necessaryfor the characterization of the protein under study. Thislimited C-terminal sequence information is often sufficientto solve problems that cannot be solved by applying anyother analytical method commonly used today.