High-throughput Method for N-terminal Sequencing of Proteins by Using Post Source Decay of MALDI Mass Spectrometry
نویسندگان
چکیده
High-throughput Method for N-terminal Sequencing of Proteins by Using Post Source Decay of MALDI Mass Spectrometry M. Yamaguchi, T. Obama, H. Kuyama, E. Ando, T. Okamura, N. Ueyama, T. Nakazawa, S. Norioka, O. Nishimura, and S. Tsunasawa Life Science Laboratory, Shimadzu Corporation, Kyoto, Japan; Graduate School of Science, Osaka University, Toyonaka, Osaka, Japan; Department of Chemistry, Nara Women’s University, Nara, Japan; Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan
منابع مشابه
High-throughput method for N-terminal sequencing of proteins by MALDI mass spectrometry.
A high-throughput method for sequencing of N termini of proteins by using postsource decay (PSD) of matrix-assisted laser desorption/ionization mass spectrometry has been developed. After a protein blotted on the PVDF membrane was successively reduced, S-alkylated, and guanidinated, its N-amino group was coupled to biotinylcysteic acid. The protein was then extracted from the membrane and diges...
متن کاملTop-Down Sequencing of Intact Proteins by MALDI In Source Decay (ISD)
Although MALDI mass spectrometry is a soft ionization technique which enables desorption of high molecular weight biomolecules exceeding several hundred kDa, it became apparent that some analyte fragmentation occurs during the MALDI process as well. The processes of fragmentation initially characterized on MALDI-TOF mass spectrometers were designated as either; (i) in-source decay (ISD), the co...
متن کاملProtein identification methods in proteomics.
A combination of high-resolution two-dimensional (2-D) polyacrylamide gel electrophoresis, highly sensitive biological mass spectrometry, and the rapidly growing protein and DNA databases has paved the way for high-throughput proteomics. This review concentrates on protein identification. We first discuss the use of protein electroblotting and Edman sequencing as tools for de novo sequencing an...
متن کاملMALDI in-source decay, from sequencing to imaging.
Matrix-assisted laser desorption/ionization (MALDI) is now a mature method allowing the identification and, more challenging, the quantification of biopolymers (proteins, nucleic acids, glycans, etc). MALDI spectra show mostly intact singly charged ions. To obtain fragments, the activation of singly charged precursors is necessary, but not efficient above 3.5 kDa, thus making MALDI MS/MS diffic...
متن کاملMatrix-assisted laser desorption/ionization time-of-flight mass spectrometry in clinical chemistry.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-Tof-MS) has recently become a popular and versatile method to analyze macromolecules from biological origin. In this paper, we will review the application of MALDI-Tof-MS in clinical chemistry and biology. MALDI-Tof-MS is used in clinical chemistry, e.g. disease markers can be identified with MALDI-MS analysis i...
متن کامل