6-Phosphogluconate dehydrogenase is a 45-kDa antigen recognized by S4D5, a monoclonal antibody specific to vanadocytes in the vanadium-rich ascidian Ascidia sydneiensis samea.
We previously prepared a monoclonal antibody, S4D5, specific to vanadocytes, vanadium-containing blood cells, in the vanadium-rich ascidian Ascidia sydneiensis samea. Here, we demonstrate that a 45-kDa antigen recognized by S4D5 is 6-phosphogluconate dehydrogenase (6-PGDH), an enzyme of the pentose phosphate pathway, based on cDNA isolation of RNA samples from blood cells of the ascidian. Western blot analysis confirmed an abundance of 6-PGDH protein in the vanadocytes and localization of 6-PGDH in the soluble extract of the blood cells. Soluble protein exhibited a correspondingly high level of 6-PGDH enzymatic activity. Ascidians are known to selectively accumulate high levels of vanadium in vanadocytes, and the highest recorded concentration of accumulated vanadium is 350 mM, which is 10(7) times the concentration in sea water. Almost all vanadium ions are reduced to the +3 oxidation state via the +4 oxidation state in vanadocytes, indicating that reducing agents must participate in the accumulation. On the other hand, vanadium ions in the +5 oxidation state are reduced to the +4 oxidation state by the presence of NADPH in vitro. Together, these observations suggest that NADPH produced in the pentose phosphate pathway may conjugate the reduction of vanadium from the +5 oxidation state through the +4 oxidation state in vanadocytes of ascidians.
Uyama
T
T
Mukaishima Marine Biological Laboratory, Faculty of Science and Laboratory of Marine Molecular Biology, Graduate School of Science, Hiroshima University, Hiroshima, 722-0073, Japan.
Kinoshita
T
T
Takahashi
H
H
Satoh
N
N
Kanamori
K
K
Michibata
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010734:Phosphogluconate Dehydrogenase / Q000235:genetics* / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D014561:Urochordata / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D014639:Vanadium / Q000378:metabolism
D000911:Antibodies, Monoclonal; D014639:Vanadium; D010734:Phosphogluconate Dehydrogenase