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J Biol Chem, Vol. 273, Issue 38, 24498-24503, September 18, 1998

Vanadium K-edge X-ray Absorption Spectroscopy Reveals Species Differences within the Same Ascidian Genera
A COMPARISON OF WHOLE BLOOD FROM ASCIDIA NIGRA AND ASCIDIA CERATODES

Patrick Frank and Keith O. Hodgson^§

From the  Department of Chemistry, Stanford University, Stanford, California 94305-5080 and the ^§ Stanford Synchrotron Radiation Laboratory, Stanford Linear Accelerator Center, Stanford University, Stanford, California 94309

Kenneth Kustin^¶, and William E. Robinson

From the ^¶ Department of Chemistry, Brandeis University, Waltham, Massachusetts 02254-9110 and the  Environmental, Coastal, and Ocean Sciences Department, University of Massachusetts, Boston, Massachusetts 02125-3393

Vanadium K-edge x-ray absorption spectroscopy (XAS) was used to examine whole blood preparations from the tunicates Ascidia nigra and Ascidia ceratodes. Each XAS spectrum exhibits a rising edge inflection near 5480 eV characteristic of vanadium(III) and an intensity maximum at 5484.0 eV. In A. ceratodes blood cells, intrinsic aquo-VSO₄⁺ complex ion is indicated by an inflection feature at 5476 eV in the first derivative of the vanadium K-edge XAS spectrum, but this feature is notably absent from the first derivative of the vanadium K-edge spectrum of blood cells from A. nigra. A strong pre-edge feature at 5468.6 eV also uniquely distinguishes the vanadium K-edge XAS spectrum of A. nigra blood cells, implying that vanadyl ion represents ~25% of the endogenous vanadium. However, the energy position of the rising edge inflection of the vanadium K-edge XAS spectrum of A. nigra (5479.5 eV) is 1 eV lower than that of A. ceratodes (5480.5 eV), the reverse of any expected shift arising from the endogenous vanadyl ion. Thus, in contrast to A. ceratodes, a significant fraction of the blood cell vanadium(III) in A. nigra is apparently in a ligation environment substantially different from that provided by water. These novel species-related differences may have taxonomic significance.

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