Hinterstoisser B, Missbichler A, Pineau B, Peschek G A
Institute of Physical Chemistry, University of Vienna, Austria.
Biochem Biophys Res Commun. 1988 Aug 15;154(3):839-46. doi: 10.1016/0006-291x(88)90216-1.
Plasma and thylakoid membranes were isolated and purified from the cyanobacterium Anacystis nidulans. Spectrophotometric examination of acetone extracts gave major absorption bands resulting from carotenoids and chlorophyll a in plasma and thylakoid membranes, respectively. Only a very small absorption peak at 663 nm was detected in acetone extracts of plasma membranes which, in contrast to the corresponding peak from thylakoid membranes, could not be extracted into n-hexane; methanol, on the other hand, was effective with both plasma and thylakoid membranes. Aqueous membrane suspensions excited at 435 nm gave strong fluorescence emission at 662 nm for plasma membranes, but only a very small one for thylakoid membranes which had been adjusted to equal absorbance at 678 nm. Excitation spectra of the 668 nm fluorescence emission peak in acetone extracts of plasma and thylakoid membranes were strikingly different from each other. Finally, high performance liquid chromatography afforded clear-cut preparative separation of the two "chlorophyll-like" pigments in plasma and thylakoid membranes, respectively, and identification by comparison with retention characteristics known from the literature, together with a pure chlorophyll a standard. Our results indicate that the highly fluorescent and polar "chlorophyll-like" pigment in plasma membranes of Anacystis is a chlorophyll precursor, viz. chlorophyllide a.
从蓝藻集胞藻中分离并纯化了质膜和类囊体膜。对丙酮提取物进行分光光度检测，结果显示质膜和类囊体膜中分别存在由类胡萝卜素和叶绿素a产生的主要吸收带。在质膜的丙酮提取物中仅检测到一个位于663nm的非常小的吸收峰，与类囊体膜相应的峰不同，该峰不能被提取到正己烷中；另一方面，甲醇对质膜和类囊体膜均有效。在435nm激发的水膜悬浮液中，质膜在662nm处发出强烈的荧光，而对于在678nm处已调节至相同吸光度的类囊体膜，仅发出非常微弱的荧光。质膜和类囊体膜的丙酮提取物中668nm荧光发射峰的激发光谱彼此明显不同。最后，高效液相色谱分别对质膜和类囊体膜中的两种“叶绿素样”色素进行了清晰的制备分离，并通过与文献中已知的保留特征以及纯叶绿素a标准品进行比较进行了鉴定。我们的结果表明，集胞藻质膜中高度荧光且极性的“叶绿素样”色素是叶绿素前体，即叶绿素酸a。
