TY - JOUR
T1 - Characteristics of structure, composition, mass spectra, and iron release from the ferritin of shark liver (Sphyrna zygaena)
AU - Huang, He Qing
AU - Xiao, Zhi Qun
AU - Chen, Xu
AU - Lin, Qing Mei
AU - CAI, Zongwei
AU - Chen, Ping
N1 - Funding Information:
This work was supported by the Natural Science Fund of China (No. 40276033), the Foundation of Fujian Provincial Natural Science in China (No. C0310006), the Graduate Scholarship Program of Xiamen University, and by the Foundation of Xiamen Natural Science in China (No.3502Z2001262).
PY - 2004/11/1
Y1 - 2004/11/1
N2 - 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 Fe3+/SZLF within the protein shell. Nature SZLF (SZLFN) consisting of holoSZLF and SZLF with unsaturated iron (SZLFUI) 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 ∼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.
AB - 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 Fe3+/SZLF within the protein shell. Nature SZLF (SZLFN) consisting of holoSZLF and SZLF with unsaturated iron (SZLFUI) 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 ∼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.
KW - AvBF
KW - bacterial ferritin of Azotobacter vinelandii
KW - Composition
KW - HLF
KW - horse spleen ferritin
KW - HSF
KW - human liver ferritin
KW - IEF
KW - isoelectric focusing
KW - Kinetics of iron release
KW - Liver ferritin
KW - liver ferritin of Sphyrna zygaena
KW - MALDI-TOF MS
KW - PSFF
KW - Shark
KW - SZLF
KW - Tumor
UR - http://www.scopus.com/inward/record.url?scp=6344263600&partnerID=8YFLogxK
U2 - 10.1016/j.bpc.2004.06.002
DO - 10.1016/j.bpc.2004.06.002
M3 - Journal article
C2 - 15501564
AN - SCOPUS:6344263600
SN - 0301-4622
VL - 111
SP - 213
EP - 222
JO - Biophysical Chemistry
JF - Biophysical Chemistry
IS - 3
ER -