Screening for carbon-bound phosphorus in marine animals by high-resolution P-31-NMR spectroscopy: coastal and hydrothermal vent invertebrates

Animals of hydrothermal vents live in a unique environment that conceivably could lead to modifications of the usual phosphorus functional groups of importance in living systems. To explore this possibility, specimens of a sea anemone (unidentified) from the TAG hydrothermal field, Mil Atlantic Ridge. the mussel Buthymodiolus N. sp. from the Mid-Atlantic Ridge, and the tubeworm Riftia puchyptila from the East Pacific Rise were analyzed for compounds containing the carbon-phosphorus bond. The analysis was based on the use of P-31-nuclear magnetic resonance, which gives signals for C-P compounds that are well separated from those of biological phosphoric acid derivatives. The animals were extracted to provide a lipid- and a water-soluble fraction, leaving an insoluble, largely proteinaceous solid residue. The lipid and residue fractions were subjected to hydrolysis to release bound forms of phosphonic acids. All fractions were analyzed by P-31-NMR. Aminophosphonic acids [primarily NH2CH2CH2PO(OH)(2) (1) and CH2NHCH2CH2PO(OH)(2) (2)1 represented the only type of C-P compound detected. These are well-known constituents of coastal invertebrates. For the mussel and sea anemone, these compounds were present in bound form in both the lipid and insoluble residue. The tube worm contained C-P material only in the insoluble residue, but in quite small amounts. The P-31-NMR method is especially valuable in being able to discriminate between compounds 1 and 2. By this technique, two coastal sea anemones (Tealia felina and Bunadosoma cavernata), previously thought to have 1 as the dominant aminophosphonic acid, were in fact found to be much richer in originally undetected 2. This compound was also detected for the first time in a mussel (Genkensia demissa), (C) 2001 Elsevier Science inc. All rights reserved.