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A manual sequence method of peptides and phosphopeptides using 4-(1'-cyanoisoindolyl)phenylisothiocyanate.


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タイトル: A manual sequence method of peptides and phosphopeptides using 4-(1'-cyanoisoindolyl)phenylisothiocyanate.
著者: Shibata, Takayuki / Wainaina, Moses N / Miyoshi, Takayuki / Kabashima, Tsutomu / Kai, Masaaki
発行日: 2011年 6月17日
出版者: Elsevier B.V.
引用: Journal of Chromatography. A, 1218(24), pp.3757-3762; 2011
抄録: A method for sequence analysis and identification of phosphoamino acids in peptides based on high performance liquid chromatography (HPLC) is described. The peptides were derivatized with an Edman type reagent, 4-(1'-cyanoisoindolyl)phenylisothiocyanate (CIPIC) and subsequently cleaved to generate stable and fluorescent 4-(1'-cyanoisoindolyl)phenylthiazolinone (CIP-TZ)-amino acids. Several experimental factors that affected derivatization on membranes were examined. Under the optimized conditions, the CIP-TZ derivatives of Try(p), Thr(p) and Ser(p) were obtained and separated from their parent amino acids with baseline resolution using an isocratic elution system. Up to the 4th residue of phosphorylated pentapeptides was successfully identified, whereas phosphoamino acid residues could not be detected by the conventional procedure using phenylisothiocyanate (PITC). The results demonstrated the potential of CIPIC as a derivatization reagent for peptide sequencing and the applicability of the method for the study and identification of phosphoamino acids in peptides.
キーワード: CIPIC / Fluorescence detection / Phosphorylated peptides / Sequencing
URI: http://hdl.handle.net/10069/25455
ISSN: 00219673
DOI: 10.1016/j.chroma.2011.04.040
PubMed ID: 21531425
権利: Copyright © 2011 Elsevier B.V. All rights reserved.
資料タイプ: Journal Article
原稿種類: author
出現コレクション:050 学術雑誌論文

引用URI : http://hdl.handle.net/10069/25455

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