褐藻光合作用色素蛋白质复合物——研究进展和问题

本文综述了近二十年来褐藻色素蛋白质复合物的研究进展,包括色素蛋白质复合物分离技术、褐藻的光系统Ｉ、光系统ＩＩ及捕光色素蛋白质复合物研究进展。并就褐藻色素蛋白质复合物分离技术中存在的问题、褐藻的特点、褐藻与其它光合生物的色素蛋白质复合物的同源性以及褐藻ＰＳＩ复合物７７Ｋ荧光发射的特点等进行了讨论     

褐藻光合作用色素_蛋白质复合物——研究进展和问题

本文综述了近二十年来褐藻色素－蛋白质复合物的研究进展,包括色素－蛋白质复合物分离、褐藻的光系统Ⅰ、光系统Ⅱ及捕光色素－蛋白质复合物研究进展。并就藻色素－蛋白质复合物分离技术中存在的问题、褐藻的特点、褐藻与其它不铪 生物的色素－蛋白质复合物的同源性以及褐藻ＰＳＩ复合物７７Ｋ荧光发射的特点等进行了讨论。     

三种不同制剂来源的叶绿素蛋白质复合物CPa的光谱特性及其组分的比较研究

用 SDS-PAGE 方法分离了菠菜叶绿体制剂、放氧光系统Ⅱ制剂和放氧光系统Ⅱ反应中心核心复合物的色素蛋白质复合物,对它们的 CPa 带进行的光谱特性的对比研究表明,在前两种制剂中 CPa 带不仅含有 Chl a 的蛋白质复合物带,它还含有少量 Chl b。且叶绿体制剂的 CPa 带中的 Chl b 含量高于放氧光系统Ⅱ制剂中的含量。此外,根据光系统Ⅱ反应中心核心复合物只有一条叶绿素蛋白质复合物带(CPa)的实验结果,我们认为光系统Ⅱ反应中心叶绿素蛋白质复合物即在 CPa 带中。但在叶绿体制剂和放氧光系统Ⅱ制剂的情况下,CPa 带还含有其它组分。     

褐藻裙带菜色素—蛋白质复合物的分离与命名

以非离子去污剂癸基－Ｎ－甲基葡萄糖胺为增溶剂,采用聚丙烯酰胺凝胶电泳技术从褐藻裙带菜（Ｕｎｄａｒｉａ ｐｉｎｎａｔｉｆｉｄａ Ｈａｒｖ．）的类囊体膜上分离到８种色素－蛋白质复合物。根据其表观分子量、光谱特性和多肽分析结果,并以高等植物菠菜（Ｓｐｉｎａｃｉａ ｏｌｅｒａｃｅａ Ｌ．）为对照,按照Ａｎｄｅｒｓｏｎ命名系统,８种色素－蛋白质复合物分别命名为ＣＰⅠ ａ、ＣＰⅠ、ＣＰａ、ＬＨＣ１、ＬＨＣ２、     

褐藻裙带菜中PSII复合物及其亚复合物的分离和特性研究

用不连续梯度蔗糖密度超离心,从经TritonX-100增溶的褐藻裙带菜类囊体膜中分离到3种色素蛋白复合物条带,分别是捕光复合物、具有光氧化活性的PSII复合物颗粒(区带II)以及PSI(区带III)。PSII颗粒经毛地黄皂苷增溶后,再次超离心分离得到3条PSII的亚复合物条带。吸收和荧光激发谱显示其中的区带II-1为墨角藻黄素-Chla/c-蛋白复合物,区带II-2为Chla/c-蛋白复合物,两者都只含20kDa多肽;而鲜绿色的区带II-3为不含捕光复合物的活性PSII核心。     

裙带菜PSⅠ复合物的荧光特异性

以褐藻裙带菜(Undaria pinnatifida)为实验材料,采用蔗糖密度梯度超速离心的方法,去污剂SDS为增溶剂(SDS:Chi=20:1,4℃增溶20 min),蔗糖密度梯度为60%、50%、40%、30%、20%、15%和10%,分离制备光系统Ⅰ(PSⅠ)复合物.结果表明,40%蔗糖层带所含色素蛋白复合物是PS I复合物.利用红藻作参照对比,光谱结果表明从裙带菜中得到的PSⅠ复合物没有730 nm的荧光峰.分析认为这是所有褐藻包括裙带菜PSⅠ复合物的荧光特异性.     

裙带菜PSⅠ复合物的荧光特异性

以褐藻裙带菜（Undaria pinnatifida）为实验材料,采用蔗糖密度梯度超速离心的方法,去污剂SDS为增溶剂（SDS:Chl＝20:1,4℃增溶20 min）,蔗糖密度梯度为60%、50%、40%、30%、20%、15%和10%,分离制备光系统Ⅰ(PSⅠ)复合物。结果表明, 40% 蔗糖层带所含色素蛋白复合物是PSⅠ复合物。利用红藻作参照对比,光谱结果表明从裙带菜中得到的PSⅠ复合物没有730 nm的荧光峰。分析认为这是所有褐藻包括裙带菜PSⅠ复合物的荧光特异性。     

褐藻裙带菜中PSⅡ复合物及其亚复合物的分离和特性研究

用不连续梯度蔗糖密度超离心,从经Triton X-100增溶的褐藻裙带菜类囊体膜中分离到3种色素蛋白复合物条带,分别是捕光复合物、具有光氧化活性的PSⅡ复合物颗粒（区带Ⅱ）以及PSⅠ（区带Ⅲ）。PSⅡ颗粒经毛地黄皂苷增溶后,再次超离心分离得到3条PSⅡ的亚复合物条带。吸收和荧光激发谱显示其中的区带Ⅱ-1为墨角藻黄素-Chl a／c-蛋白复合物,区带Ⅱ-2为Chl a／c-蛋白复合物,两者都只含20kDa多肽;而鲜绿色的区带Ⅱ-3为不含捕光复合物的活性PSⅡ核心。     

光逆境对叶绿体叶绿素蛋白质复合物的影响

研究了强光照射对菠菜叶绿体的叶绿素蛋白质复合物及一些光合特性的影响。实验结果表明,随着强光照射时间的延长,首先,属光系统Ⅱ核心的叶绿素蛋白质复合物的CPa带明显减少了,进而属LHCII的寡聚体和二聚体的带有了不同程度的降低,最后,包括光系统I在内的叶绿素蛋白质复合物带大部分被分解了。结果还表明,当光逆境还未使叶绿素蛋白质复合物发生明显变化时,代表光系统Ⅱ活性的Fv/Fo值及DCIP光还原活性就已显著地降低了。     

植物光系统II的组份和辅因子在其组装过程中的作用和调节

光系统II(photosystem II,PSII)是光合作用光反应过程重要的光合膜蛋白复合体。它是由大约25个不同蛋白质复合物及其辅因子组成的色素蛋白复合体。由于PSII结构的复杂性,PSII的组装是多步骤的,并得到辅因子和调控蛋白的协助。重点讨论PSII组份色素、小亚基、外周蛋白和保守因子在其组装过程中的作用和调节机制,并介绍了蓝细菌和植物叶绿体中的一些特殊蛋白质调控因子。     

管藻目绿藻叶绿素蛋白复合物特性及比较研究

By mild PAGE method, 11, 11, 7 and 9 chlorophyll-protein complexes were isolated from two species of siphonous green algae (Codium fragile (Sur.) Hariot and Bryopsis corticulans Setch.), green alga (Ulothrix flacca (Dillw.) Thur.), and spinach (Spinacia oleracea Mill.), respectively. Apparent molecular weights, Chl a/b ratios, distribution of chlorophyll, absorption spectra, low temperature fluorescence spectra of these complexes were determined, and compared with one another. PSⅠ complexes of two siphonous green algae are larger in apparent molecular weight because of the attachment of relative highly aggregated LHCⅠ. Four isolated light-harvesting complexes of PSⅡ are all siphonaxanthin-Chl a/b-protein complexes, and they are not monomers and oligomers like those in higher plants. Especially, the absence of 730 nm fluorescence in PSⅠ complexes indicates a distinct structure and energy transfer pattern.     

The Chlorophyll-Protein Complexes Resolved from Ultrasonically Broken Thylakoid Memb-ranes of Blue-Green Algae

When the thylakoid membranes of blue-green algae were broken by ultrasonic vibrations and subjected to polyacrylamide gel electrophoresis at 4℃, six green zones were resolved. They were designated as CPIa, CPlb, CPI; CPal, CPa2, and FC. The absorption spectrum of CPI had a red maximum at 674 nm and a peak in the blue at 435 nm. It was identified as PS chlorophyll a-protein Complex, but was contaminated with minor PSⅡ which was implied by the appearance of fluorescence emission peak at 680 nm besides the main one at 725 nm at 77 K. The spectral properties of CPIa and CPlb were similar to that of CPl. The absorption spectra of CPa1 and CPa2 were similar, both having red maxima at 667 nm and peaks in the blue at 431.5 nm. Their fluorescence emission had the same peaks at 684 nm at 77 K indicating that they belonged to PSⅡ. It was recognized that CPal of 47 kD is the reaction center complex of photosystem Ⅱ and CPa2 of 40 kD is the internal antenna complex of photosystem Ⅱ. The spectral characteristics of the chlorophyll-protein complexes resolved by ultrasonic method were similar to those of the same complexes resolved by SDS solubilization, except the absorbance positions of CPa1 and CPa2 in the blue peak and the red one which shifted to blue about 3–5 nm. It was calculated that in thylakoid membranes of blue-green algae 40.93% chlorophyll was in PSⅠ, while 38.78% of chlorophyll in PSⅡ. The difference of chlorophyll contents between PSⅠ and PSⅡ was only 2.15%. Concerning the fact that minor PSⅡ compound remained in the part of PSⅠ zones, it might be concluded that the distribution of chlorophyll between PSⅠ and PSⅡ in blue-green algae was equal. This result was in agreement with the hypothesis that PSⅠ and PSⅡ operates in series in photosynthetic electron transport.     

Chlorophyll-Protein Complexes of Blue-Green Algae Isolated by a New Gel Electrophoresis System with High Resolving Power

At least 13 chlorophyll bands from the thylakoid membranes of blue-green algae could be clearly resolved by SDS-PAGE employing a new improved procedure. They were designated as CPIa, CPIb, CPIc, CPId, CPIe, CPIf, CPIg, CHIh, CPal, CPa2, CPa3, CPa4 and FC. 8 chlorophyll-protein complexes, CPIa-CPIh, had the same absorption spectrum at 676 nm in the red and 436 nm in the blue region. They belonged to the chlorophyll-protein complexes of PS Ⅰ. 4 chlorophyll-protein complexes, CPal-CPa4, had a red absorption peak at 670­672 nm and a blue one at 436 nm. Their fluorescence emission peak at 77K was at 685 nm. They were chlorophyll-protein complexes of PS Ⅱ.     

Effects of Water Stress on Chlorophyll-Protein Complexes in Wheat Chloroplasts

The excitation energy transfer from light harvesting chlorophyll protein complexes to PS Ⅱ was inhibited under water stress. The contents of iriternal antennae chlorophyll-protein complexes of PS Ⅱ (CPa), light harvesting chlorophyll-protein complexes of PS Ⅱ (LHC Ⅱ ), light harvesting chlorophyll-protein of PS Ⅰ (LHC Ⅰ ) and chlorophyll a protein complex of reaction center of PS Ⅰ were decreased by water stress. The decrease of chlorophyll-protein complexes of PS Ⅱ was greater than that of PS Ⅰ . It was indicated that the amount of 25 kD polypeptide of LHC Ⅱ in particular, as well as that of 43 and 47 kD polypeptides of CPa, and 21 kD polypeptide of LHC Ⅰ , were reduced by water stress.     

The Regulation and Distribution of LHC-I and LHCP2 between the Photosystem I and Photosystem II

Evidences were provided in this paper that the relative distribution of chl-protein complexes of PSⅠ and PSⅡ could be regulated by Mg2+. addition of Mg2+ led to decrease in the amount of chl-protein complexes of PSⅠ and increase in the amount of chl-protein in complexes of PSⅡ. There was no effect of Mg2+ on the spectral property of LHCP1, but the addition of Mg2+ could change the spectral property of LHCP2 so that it became similar to that of the LHC-Ⅰ. CPIa2 was a complex of reaction centre of PSⅠ and LHC-I. LHC-I might be contacted specially with LHCP2 in chloroplast membranes. Addition of Mg2+ probably cansed the motion of LHC-I from PSⅠ to PSⅡ and became more closely connected with LHCP2. The relative amount of CPIa2, CPIa1, LHCP1 and LHCP2 in chloroplast membranes could be regulated by different light intensity. There were more CPIa2, LHCP1 and less LHCP2 in chloroplast membranes from the shade plant Malaxis monophyllos and sunflower grown under weak light, both of them lacked equally CPIa1. There were less CPIa2, LHCP1 and more LHCP2 in the sun plant spinach and sunflower grown under strong light, and they possessed equally CPIa1 chl-protein complexes. It is suggested that LHCP1 and LHCP2 are different light-harvesting Chl-protein complexes. The LHC-I and LHCP2 are mobile light-harvesting chl-protein complexes and shuttle back and forth between PSⅠ and PSⅡ They play an important role in the regulation and distribution of excitation energy between the two photosystems.     

The Possible Mechanism of Photoinhibition of Purified PSI Complexes

The protecting effect of histidine on the photodamage of pigments and proteins of the isolated PSⅠ particles from the chloroplast of Spinacia oleracea L. during the strong illumination (2 300 μmol·m-2·s-1) was studied by spectroscopy and SDS-PAGE. The absorbance of PSⅠ particles decreased during the strong illumination treatment, but the decrease would be slowed down in the presence of externally added histidine after 30 min illumination. The decrease of CD （circular dichroism）signal intensities of PSⅠ particles also was slowed down by the added histidine after about 10 min illumination. The retarded protecting effect of the added histidine on the photobleaching of pigments of PSⅠ complexes implied that the mechanisms of photoinhibition of isolated PSⅠ complexes are different from early stage to later stage during the strong illumination treatment. In addition, the added histidine suppressed the decrease of 77 K fluorescence yield of PSⅠ particles during the illumination. SDS-PAGE showed that the added histidine not only protected the reaction center proteins of PSⅠ particles, but also protected other subunits of PSⅠ particles from degradation.     

Isolation and Nomenclature of Pigment-Protein Complexes from the Brown Alga (Undaria pinnatifida Harv.)

Eight kinds of pigment-protein complexes were resolved from the thylakoid membrane of the brown alga （Undaria pinnatifida Harv.) by using non-ionic detergent decanoyl-N-methylglucamide and PAGE technique. According to the apparent molecular weights, spectra characteristics, polypeptide compositions and referring to the higher plant spinach, eight pigment-protein complexes were named under Anderson′s terminology system as CPⅠa, CPⅠ, CPa, LHC1, LHC2, LHC3, LHC4, LHC5.     

The supramolecular architecture, function, and regulation of thylakoid membranes in red algae: an overview

Red algae are a group of eukaryotic photosynthetic organisms. Phycobilisomes (PBSs), which are composed of various types of phycobiliproteins and linker polypeptides, are the main light-harvesting antennae in red algae, as in cyanobacteria. Two morphological types of PBSs, hemispherical- and hemidiscoidal-shaped, are found in different red algae species. PBSs harvest solar energy and efficiently transfer it to photosystem II (PS II) and finally to photosystem I (PS I). The PS I of red algae uses light-harvesting complex of PS I (LHC I) as a light-harvesting antennae, which is phylogenetically related to the LHC I found in higher plants. PBSs, PS II, and PS I are all distributed throughout the entire thylakoid membrane, a pattern that is different from the one found in higher plants. Photosynthesis processes, especially those of the light reactions, are carried out by the supramolecular complexes located in/on the thylakoid membranes. Here, the supramolecular architecture, function and regulation of thylakoid membranes in red algal are reviewed.