Fluorometric determination of thiol and disulfide groups in protein using N-(9-acridinyl)maleimide.

• HATAKEYAMA Eiko

  Laboratory of Food Hygiene, Tohoku Fukushi University

• MATSUMOTO Norikazu

  Laboratory of Food Hygiene, Tohoku Fukushi University

• OCHI Takeo

  Laboratory of Food Hygiene, Tohoku Fukushi University

• SUZUKI Tateo

  Laboratory of Wholesomeness Evaluation, National Food Research Institute

• OHRUI Hiroshi

  Laboratory of Analytical Chemistry, Faculty of Agriculture, Tohoku University

• MEGURO Hiroshi

  Laboratory of Analytical Chemistry, Faculty of Agriculture, Tohoku University

抄録

A sensitive fluorometric method has been developed to determine the thiol and disulfide groups in proteins using N-(9-acridiny )maleimide as a fuorogenic reagent. Proteins are hydrolyzed with subtilopeptidase (Carlsberg type) in order to eliminate the steric hindrance. Then, thiols are determined by the reaction of N-(9-acridinyl)maleimide with the hydro ysate. Disulfide can be determined using the same hydro ysate after the derivatization into thins by<br>electroytical reduction. The fluorescence intensities have shown linear relations between the thin and disulfide contents at nmol/ml levels in the mode proteins. The sensitivity is 1000-times higher than conventional colorimetry with 5, 5′-dithiobis(2-nitrobenzoate).

収録刊行物

• Analytical Sciences

  Analytical Sciences 5 (6), 657-661, 1989

  社団法人 日本分析化学会

キーワード

• Fluorometry
• thiol
• disulfide
• protein
• <i>N</i>-(9-acridiny)maleimide

詳細情報

• CRID: 1390001204257493632
• NII論文ID: 130003528883
• DOI: 10.2116/analsci.5.657
• COI: 1:CAS:528:DyaK3cXhslWntb8%3D
• ISSN: 13482246
• ISSN: 09106340
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.5.657.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.5.657/fulltext.html
• Web Site: http://www.jstage.jst.go.jp/article/analsci1985/5/6/5_6_657/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • Crossref
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可