Direct Detection of Mercury-Bound Metalloproteins (metallothionein and Cu, Zn-Superoxide Dismutase) Using a Combination of Gel Electrophoresis and One Dimensional Synchrotron Radiation X-Ray Fluorescence Analysis
作者:
S. HOMMA-TAKEDA,
M. SHINYASHIKI,
I. NAKAI,
C. TOHYAMA,
Y. KUMAGAI,
N. SHIMOJO,
期刊:
Analytical Letters
(Taylor Available online 1996)
卷期:
Volume 29,
issue 4
页码: 601-611
ISSN:0003-2719
年代: 1996
DOI:10.1080/00032719608000424
出版商: Taylor & Francis Group
关键词: electrophoresis;synchrotron radiation X-ray fluorescence analysis;mercury;metallothionein;Cu, Zn-superoxide dismutase
数据来源: Taylor
摘要:
Metallothionein-II (MT-II) and Cu, Zn-superoxide dismutase (Cu, Zn-SOD) interacted with mercury were detected by a new method utilizing isoelectric focusing-agarose or -polyacrylamide gel electrophoresis (IEF-AGE or IEF-PAGE) and nondestructive one-dimensional synchrotron radiation X-ray fluorescence (SR-XRF) analysis. When MT-II reacted with mercuric chloride, an obvious change of isoelectric point (pI = 3.7 - 4.7) for the intact form to alkaline pI (9.4) was observed. This marked migration of MT-II by the metal was blocked by addition of glutathione, suggesting that sulfhydryl functions participate in the pI variation. In contrast, interaction of Cu, Zn-SOD with mercury did not cause any changes of its pI although the metal bound tightly to Cu, Zn-SOD after electrophoresis; however, the enzyme activity was drastically suppressed. These observations indicate that combination of electrophoresis with SR-XRF analysis is an useful technique for detecting structural or functional alteration of protein attributable to the binding of the mercury.
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