Selective isolation of N-terminal peptides from proteins and theirde novosequencing by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry without regard to unblocking or blocking of N-terminal amino acids
| العنوان: | Selective isolation of N-terminal peptides from proteins and theirde novosequencing by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry without regard to unblocking or blocking of N-terminal amino acids |
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| المؤلفون: | Susumu Tsunasawa, Hiroki Kuyama, Norikazu Ueyama, Minoru Yamaguchi, Daisuke Nakayama, Keisuke Shima, Taka-aki Okamura, Shigemi Norioka, Eiji Ando, Takashi Nakazawa, Osamu Nishimura |
| المصدر: | Rapid Communications in Mass Spectrometry. 22:3313-3319 |
| بيانات النشر: | Wiley, 2008. |
| سنة النشر: | 2008 |
| مصطلحات موضوعية: | Protein mass spectrometry, Sequence analysis, medicine.medical_treatment, Mass spectrometry, Sample preparation in mass spectrometry, Analytical Chemistry, chemistry.chemical_compound, Cell Line, Tumor, medicine, Humans, Amino Acids, Databases, Protein, Spectroscopy, chemistry.chemical_classification, Performic acid, Protease, Chromatography, biology, Organic Chemistry, Proteins, Amino acid, chemistry, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, biology.protein, Electrophoresis, Polyacrylamide Gel, Indicators and Reagents, Peptides, Avidin |
| الوصف: | We have developed a new method to determine the N-terminal amino acid sequences of proteins, regardless of whether their N-termini are modified. This method consists of the following five steps: (1) reduction, S-alkylation and guanidination for targeted proteins; (2) coupling of sulfo-NHS-SS-biotin to N(alpha)-amino groups of proteins; (3) digestion of the modified proteins by an appropriate protease followed by oxidation with performic acid; (4) specific isolation of N-terminal peptides from digests using DITC resins; (5) de novo sequence analysis of the N-terminal peptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) using the CAF (chemically assisted fragmentation) method or tandem mass spectrometric (MS/MS) analysis according to unblocked or blocked peptides, respectively. By employing DITC resins instead of avidin resins used in our previous method (Yamaguchi et al., Rapid Commun. Mass Spectrom. 2007; 21: 3329), it has been possible to isolate selectively N-terminal peptides from proteins regardless of modification of N-terminal amino acids. Here we propose a universal method for N-terminal sequence analysis of proteins. |
| تدمد: | 1097-0231 0951-4198 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9cd339b4da9c284330e4ba922dd953e5 https://doi.org/10.1002/rcm.3735 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....9cd339b4da9c284330e4ba922dd953e5 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10970231 09514198 |
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