کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10532931 | 961802 | 2010 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A spectrophotometric method for the determination of zinc, copper, and cobalt ions in metalloproteins using Zincon
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کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Zincon (2-carboxy-2â²-hydroxy-5â²-sulfoformazylbenzene) has long been known as an excellent colorimetric reagent for the detection of zinc and copper ions in aqueous solution. To extend the chelator's versatility to the quantification of metal ions in metalloproteins, the spectral properties of Zincon and its complexes with Zn2+, Cu2+, and Co2+ were investigated in the presence of guanidine hydrochloride and urea, two common denaturants used to labilize metal ions in proteins. These studies revealed the detection of metals to be generally more sensitive with urea. In addition, pH profiles recorded for these metals indicated the optimal pH for complex formation and stability to be 9.0. As a consequence, an optimized method that allows the facile determination of Zn2+, Cu2+, and Co2+ with detection limits in the high nanomolar range is presented. Furthermore, a simple two-step procedure for the quantification of both Zn2+ and Cu2+ within the same sample is described. Using the prototypical Cu2+/Zn2+-protein superoxide dismutase as an example, the effectiveness of this method of dual metal quantification in metalloproteins is demonstrated. Thus, the spectrophotometric determination of metal ions with Zincon can be exploited as a rapid and inexpensive means of assessing the metal contents of zinc-, copper-, cobalt-, and zinc/copper-containing proteins.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 397, Issue 2, 15 February 2010, Pages 218-226
Journal: Analytical Biochemistry - Volume 397, Issue 2, 15 February 2010, Pages 218-226
نویسندگان
Crystal E. Säbel, Joseph M. Neureuther, Stefan Siemann,