193 nm Ultraviolet Photodissociation of Imidazolinylated Lys-N Peptides for De Novo Sequencing

详细信息    查看全文

• 作者：Michelle R. Robinson ; James A. Madsen ; Jennifer S. Brodbelt
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：March 6, 2012
• 年：2012
• 卷：84
• 期：5
• 页码：2433-2439
• 全文大小：522K
• 年卷期：v.84,no.5(March 6, 2012)
• ISSN：1520-6882

文摘

The goal of many MS/MS de novo sequencing strategies is to generate a single product ion series that can be used to determine the precursor ion sequence. Most methods fall short of achieving such simplified spectra, and the presence of additional ion series impede peptide identification. The present study aims to solve the problem of confounding ion series by enhancing the formation of 鈥済olden鈥?sets of a, b, and c ions for sequencing. Taking advantage of the characteristic mass differences between the golden ions allows N-terminal fragments to be readily identified while other ion series are excluded. By combining the use of Lys-N, an alternate protease, to produce peptides with lysine residues at each N-terminus with subsequent imidazolinylation of the 蔚-amino group of each lysine, peptides with highly basic sites localized at each N-terminus are generated. Subsequent MS/MS analysis by using 193 nm ultraviolet photodissociation (UVPD) results in enhanced formation of the diagnostic golden pairs and golden triplets that are ideal for de novo sequencing.