Bruno J G, Collard S B, Andrews A R
Applied Research Associates, Tyndall Air Force Base, FL 32403, USA.
J Biolumin Chemilumin. 1997 May-Jun;12(3):155-64. doi: 10.1002/(SICI)1099-1271(199705/06)12:3<155::AID-BIO442>3.0.CO;2-T.
Electrophoresis, size exclusion chromatography, fluorescence, and electrochemiluminescence (ECL) data obtained from the cell-rich perivisceral fluid ('blood') of the tunicate, Molgula occidentalis, suggests that a yellowish protein or protein subunit of < 6.5 kDa is probably responsible for the low level intrinsic ECL reported previously. Variable potential ECL scans and spectrofluorometric analyses directly indicated that chlorophylls from ingested phytoplankton were not significant contaminants of M. occidentalis blood samples. Chlorophylls were also examined indirectly from monocultures of dinoflagellate and diatom species to determine their ECL levels. While ECL was observed in these cultures, high concentrations (10(6) organisms/mL) of dinoflagellates or diatoms were needed to produce ECL levels comparable to those observed for M. occidentalis blood. Additional work with two synthetic tunichrome isomers indicated 10-fold increases in ECL when the '2,3,4-tunichrome' was reacted with Tl+ in 1:1 and 1:2 metal ion to ligand molar ratios. The '3,4,5-tunichrome' isomer exhibited a titration curve with Hg2+ suggestive of the existence of at least two Hg2+ binding sites and a 9- to 10-fold increase in ECL output.
从被囊动物西方海鞘富含细胞的围脏液（“血液”）中获得的电泳、尺寸排阻色谱、荧光和电化学发光（ECL）数据表明，一种分子量小于6.5 kDa的淡黄色蛋白质或蛋白质亚基可能是先前报道的低水平固有ECL的原因。可变电位ECL扫描和荧光光谱分析直接表明，摄入的浮游植物中的叶绿素不是西方海鞘血液样本的重要污染物。还从甲藻和硅藻物种的单培养物中间接检测了叶绿素，以确定它们的ECL水平。虽然在这些培养物中观察到了ECL，但需要高浓度（10⁶个生物体/毫升）的甲藻或硅藻才能产生与西方海鞘血液中观察到的ECL水平相当的ECL。对两种合成被囊色素异构体的进一步研究表明，当“2,3,4-被囊色素”与Tl⁺以1:1和1:2的金属离子与配体摩尔比反应时，ECL增加了10倍。“3,4,5-被囊色素”异构体与Hg²⁺的滴定曲线表明至少存在两个Hg²⁺结合位点，并且ECL输出增加了9至10倍。
