Characteristics of structure, composition, mass spectra, and iron release from the ferritin of shark liver (Sphyrna zygaena)

被引:18
作者
Huang, HQ [1 ]
Xiao, ZQ
Chen, X
Lin, QM
Cai, ZW
Chen, P
机构
[1] Xiamen Univ, Dept Biol, Sch Life Sci, Key Lab Chem Biol Fujian Province, Xiamen 361005, Peoples R China
[2] Xiamen Univ, Minist Educ, Key Lab Marine Environm Sci, Xiamen 361005, Peoples R China
[3] Xiamen Univ, Key Lab Marine Environm Sci, Minist Educ, Xiamen 361005, Peoples R China
[4] Xiamen Univ, Key Lab MOE Cell Biol & Tumor Cell Engn, Xiamen 361005, Peoples R China
[5] Hong Kong Baptist Univ, Dept Chem, Kowloon, Hong Kong, Peoples R China
基金
中国国家自然科学基金;
关键词
liver ferritin; shark; MALDI-TOF MS; kinetics of iron release; composition; tumor;
D O I
10.1016/j.bpc.2004.06.002
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The ferritin consists of a protein shell constructed of 24 subunits and an iron core. The liver ferritin of Sphyrna zygaena (SZLF) purified by column chromatography is a protein composed of eight ferritins containing varying iron numbers ranging from 400 +/- 20 Fe3+/SZLF to 1890 +/- 20 F3+/SZLF within the protein shell. Nature SZLF (SZLF(N)) consisting of holoSZLF and SZLF with unsaturated iron (SZLF(UL)) to have been purified with polyacrylamide gel electrophoresis (PAGE) exhibited five ferritin bands with different pI values ranging from 4.0 to 7.0 in the gel slab of isoelectric focusing (IEF). HoloSZLF purified by PAGE (SZLFE) not only had 1890 +/- 20 Fe3+/SZLFE but also showed an identical size of iron core observed by transmission electron microscopy (TEM). Molecular weight of similar to 21 kDa for SZLFE subunit was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Four peaks of molecular ions at mass/charge (m/z) ratios of 10611.07, 21066.52, 41993.16, and 63555.64 that come from the SZLFE were determined by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF MS), which were identified as molecular ions of the ferritin subunit (M+) and its polymers, namely, [M](2+), [M](+), [2M](+), and [3M](+), respectively. Both SZLFE and a crude extract from shark liver of S. zygaena showed similar kinetic characteristics of complete iron release with biphasic behavior. In addition, a combined technique of visible spectrometry and column chromatography was used for studying ratio of phosphate to Fe3+ within the SZLFE core. Interestingly, this ratio maintained invariable even after the iron release, which differed from that of other mammal ferritins. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:213 / 222
页数:10
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