- Autor(in)
- Seitenbereich
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129 - 146
- Zusammenfsg.
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A method is described to prepare vital cells from tentacle tissue of the sea anemone Anemonia sulcata. The isolation includes the dispersion of tentacle tissue with papain and the separation/purification of the cells by density centrifugation with self-generating Percoll density gradients. Transport of amino acids into freshly isolated cells from tissue of a free-living marine invertebrate species has been studied for the first time. Amino acid influx is composed of a saturable and a non-saturable component and is linear for at least 3 min. For the saturable transport component the apparent affinity is high (K-tapp. = 1.8 - 3.7 mu M) and the transport capacity is low (J(max) = 32 pmoles x mg protein(-1) x min(-1)). The uptake of L-leucine is temperature dependent. Q(10) values range between 1.8 and 2.8. Above 10 degrees C, the energy of activation (E(a)) was 43 kJ/Mol, below 10 degrees C, E(a) was 67 kJ/Mol. At 19 parts per thousand salinity, the uptake was reduced by 70% compared to 37.5 parts per thousand 2,4-Dinitrophenol (2 mM) reduced L-leucine uptake by 65% compared to the controls. Cross-inhibition experiments suggest the existence of at least nine separate transport systems for amino acids with different specificities.
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