Immunochemistry on cryptomonad biliproteins

A survey is made of the immunochemical behavior of four of the six known types of cryptomonad biliproteins: phycocyanins 612 and 645 and phycoerythrins 545 and 566. They were compared both among themselves and to selected biliproteins isolated from blue-green and red algae. All the cryptomonad biliproteins were shown to be closely related to each other by Ouchterlony double diffusion technics. An antigenic relationship among all the cryptomonad biliproteins and B-phycoerythrin (red alga) and C-phycoerythrin (blue-green alga) was established. Only a very marginal cross-reactivity was found between C-phycocyanin (blue-green algae) and the cryptomonad biliproteins. These results suggest a common ancestor for the photosynthetic units of all three biliprotein-containing phyla.     

Purification and N-terminal analyses of algal biliproteins

1. R-, B- and C-phycoerythrins and R- and C-phycocyanins were isolated and purified on a preparative scale by calcium phosphate chromatography, ammonium sulphate fractionation and crystallization. 2. The N-terminal residues of these biliproteins were analysed. Methionine is the only N-terminal residue of all the phycoerythrins, there being about 14 N-terminal residues per molecule of R- and B-phycoerythrins (mol.wt. 290000) and about 8 per molecule of C-phycoerythrin (mol.wt. 226000). Threonine (1 residue) is N-terminal in C-phycocyanin (mol.wt. 138000), and both threonine (about 1·3 residues) and methionine (5 residues) are N-terminal in R-phycocyanin (mol.wt. 273000). 3. Results suggest that the apoproteins of the various phycoerythrins are closely related, whereas C-phycocyanin has quite a different gross structure, and that R-phycocyanin contains two types of sub-unit, one related to C-phycocyanin and the other to the phycoerythrins.     

The native forms of the phycobilin chromophores of algal biliproteins. A clarification.

Pigments released from phycoerythrins and phycocyanins by treatment with hot methanol are currently regarded as equivalent to the native chromophores phycoerythrobilin and phycocyanobilin. However, evidence presented here confirms the original view of O'Carra & O'hEocha [(1966 Phytochemistry 5, 993-997] that these methanol-released pigments are artefacts differing in their chromophoric conjugated systems from the native protein-bound prosthetic groups. By contrast, the native spectral properties are retained in pigments released by careful acid treatment of the biliproteins and these acid-released phycobilins, rather than the methanol-released pigments, are therefore regarded as the protein-free forms of the native chromophores. The conclusion reached by Chapman, Cole & Siegelman [(1968) J. Am. Chem. Soc. 89, 3643-3645], that all the algal biliproteins contain only phycoerythrobilin and phycocyanobilin, is shown to be incorrect. The identification of a urobilinoid chromophore, phycourobilin, accompanying phycoerythrobilin in B- and R- phycoerythrins is confirmed and supported by more extensive evidence. The cryptomonad phycocyanins are shown to contain a phycobilin chromophore accompanying phycocyanobilin. This further phycobilin has the spectral properties of the class of bilins known as violins and the provisional name "cryptoviolin" is proposed pending elucidation of its structure.     

The immunologically conserved phycobilisome-thylakoid linker polypeptide

We have isolated phycobilisomes from two classes of red algae, several subdivisions of the cyanobacteria, and the cyanelles of Cyanophora paradoxa. In addition to the major light harvesting biliproteins, these phycobilisomes also contain several other polypeptides, the largest of which ranges from 75 to 120 kilodaltons in the different species surveyed. This protein, previously isolated and characterized from three species, was shown to be the final emitter of excitation energy in phycobilisomes and is also thought to be involved in the attachment of the phycobilisomes to the thylakoid membrane. We have obtained polyclonal antibodies to the 95 kilodalton polypeptide isolated from phycobilisomes of the cyanobacterium, Nostoc sp. This protein shares no common antigenic determinants with either the α or β subunits of allophycocyanin, or any of the other biliproteins, as determined by the sensitive Western immunoblotting technique. However, this antiserum cross-reacts with the highest molecular weight polypeptide of all the rhodophytan and cyanobacterial phycobilisomes tested. That these proteins are immunologically related, but are unrelated to other biliproteins, is reminiscent of previous immunological studies of biliproteins which showed that while the three major spectroscopically distinct classes of biliproteins (phycoerythrin, phycocyanin, and allophycocyanin) shared no common antigenic determinants, there was a strong antigenic determinant to specific biliprotein classes which crossed taxonomic divisions.     

Disc gel electrophoresis of biliproteins

Phycobilin proteins from eleven genera of red algae (Rhodophyta)were examined by polyacrylamide disc gel electrophoresis. Therelative electrophoretic mobility of the biliproteins indicatedmuch greater heterogeneity than previously reported. Comparisonsof protein homologies may be used in describing systematic andphylogenetic affinities of the Rhodophyta. (Received July 20, 1971; )     

Characterization of the biliproteins of Gloeobacter violaceus chromophore content of a cyanobacterial phycoerythrin carrying phycourobilin chromophore

The biliproteins of the unicellular, thylakoid-less cyanobacterium Gleobacter violaceus were resolved by chromatography on hydroxylapatite and DEAE-cellulose into five components: phycoerythrin I and II, phycocyanin I and II, and allophycocyanin. Allophycocyanin B was not detected. Three of these components, phycoerythrin II, phycocyanin II, and allophycocyanin, were purified to homogeneity. Phycoerythrin II crystallized as hexagonal prisms. G. violaceus allophycocyanin crystallized as thin plates; unter similar conditions other cyanobacterial allophycocyanins crystallize as needles. The biliproteins in the phycoerythrin I and phycocyanin I components were present in polydisperse, high molecular weight aggregates, which may represent incompletely dissociated substructures of the phycobilisome.Both phycoerythrin components from G. violaceus carry phycoerythrobilin and phycourbilin groups in the ratio of 6:1. Separation of the and subunits of these biliproteins revealed that the phycoerythrobilins were equally distributed between the two subunits, and that the subunit alone carried the phycourobilin. These phycoerythrins are the first cyanobacterial phycobiliproteins found to carry a phycourobilin prosthetic group.Abbreviations used PE poycoerythrin - PC phycocyanin - AP allophycocyanin - SDS sodium dodecyl sulfate - PAGE polyacrylamide gel electrophoresis - B Bangiophycean - R Rhodophytan - C Cyanobacterial     

The horizontal distribution of the benthic algal flora in the Patos Lagoon estuary, Brazil, in relation to salinity, substratum and wave exposure

Monthly collections of benthic algae were made during 1 yr at 11 stations between the mouth and the salt water limits of the Patos Lagoon estuary, Brazil. A total of 94 taxa of algae were recorded, including 40 Cyanophyta, 26 Chlorophyta, 1 Xanthophyta, 3 Phaeophyta, and 24 Rhodophyta. Although the number of Rhodophyta and Phaeophyta decreased towards lower salinities, due to local salinity patterns, species substitution among the Chlorophyta and increase of Cyanophyta in the estuary, the species number remained approximately the same between the mouth and the upper saline reaches of this estuary.

The distributional limits of 64 species in the flora resulted in the formation of three algal groups of negative correlation. The horizontal distribution of each group of species was related to type of substratum, wave exposure or salinity, or a combination of these factors. The individual evaluation of the factors demonstrated that, apart from salinity, substratum and degree of exposure are of primary importance for the horizontal distribution of a large part of the algal flora in this estuary.     

Characterization of tidal pool algae in the Mexican Tropical Pacific coast

Tidal pools in the Mexican Tropical Pacific coast have received relatively little attention in spite of their considerable richness in species and wide distribution in the region.This paper presents the first characterization of the algal flora of this region. It analyzes the number and composition of species of the tidal pools of six localities with regard to geographical distribution and its seasonal variations as well as tidal level. 97 species are reported, 25 Chlorophyta, 23 Phaeophyta, 34 Rhodophyta and 15 Cyanophyta.Of that total of species, 63% were found in one locality, 23.7% in two, 11.3% in three and 1 % in 4 or 5 localities. Not one species was common to all of the pools.The highest number of species was found on pools of the middle and low intertidal where the Chlorophyta, Rhodophyta and Phaeophyta were the most abundant algae. Cyanophyta was more common in the supralittoral and high intertidal pools.     

Rhodosorusmarinus中澡红蛋白的纯化及其性质的研究

从Ｒｈｏｄｏｓｏｒｕｓｍａｒｉｎｕｓ中提取了藻红蛋白,通过改进纯化方法,得到了三种电泳纯的藻红蛋白：Ｂ－型藻红蛋白１,Ｂ－型藻红蛋白２和ｂ－型藻红蛋白（以下简称Ｂ—ＰＥ１,Ｂ—ＰＥ２和ｂ－ＰＥ）。分别测定这三种藻红蛋白聚合体和亚单位的分子量。测定了它们的可见光的吸收光谱和荧光光谱。两种Ｂ—ＰＥ的可见光的吸收光谱比ｂ—ＰＥ的多一个４９８ｎｍ的吸收峰。三种藻红蛋白的氨基酸组成以酸性氨基酸和疏水性氨基酸为主,所以经等电聚焦测定的它们的等电点都是偏酸性的。     

Structural studies on cryptomonad biliprotein subunits. Two different alpha-subunits in Chroomonas phycocyanin-645 and Cryptomonas phycoerythrin-545

The N-terminal amino-acid sequences of two green alpha-subunit fractions from Chroomonas phycocyanin-645 and from two violet alpha-subunit fractions from Cryptomonas phycoerythrin-545 reveal that these cryptomonad biliproteins each contain two different alpha-subunits. The chromophore binding sites at the cysteine residues in positions 18 or 19 are homologous to the chromophore binding site at cysteine position 84 in cyanobacterial biliproteins. The sequence homologies of the beta-subunits to cyanobacterial biliproteins are higher than those of the alpha-subunits. Cryptomonas phycoerythrin-545 alpha-subunits contain a gamma-hydroxylysine residue at the fourth position of the polypeptide chains. 50%-75% of the total sequence of the alpha-subunits was determined by N-terminal amino-acid sequence analysis. The alpha-subunits of the Cryptomonad biliproteins are smaller than the alpha-subunits of the cyanobacterial biliproteins. Comparing sequence homologies we found 60 amino-acid residues less at the N-terminus of Cryptomonad biliproteins than in cyanobacterial biliproteins.     

Comparison of phycoerythrins (542, 566 nm) from cryptophycean algae

Summary The phycoerythrins from Rhodomonas sp. strain 3-C and Cryptomonas ovata var. palustris were purified and partially characterized. The phycoerythrin from Rhodomonas had a single visible absorption maximum at 542 nm with a shoulder at approximately 562 nm and is, therefore, representative of cryptophyte type I phycoerythrin. The phycoerythrin from C. ovata var. palustris had a single absorption maximum at 566 nm and is, therefore, representative of cryptophyte type III phycoerythrin. Calibrated gel filtration chromatography showed that both of these phycoerythrins have a native molecular weight of 30 800 daltons. Calibrated sodium dodecyl sulfate gel electrophoresis demonstrated that both pigments were composed of two subunits with apparent molecular weights of 17 700 and 11 000 daltons. On polyacrylamide gel electrofocusing both these phycoerythrins had an isoionic point of 4.90.     

Epiphytic algae from Bajo Pepito, Isla Mujeres, Quintana Roo, Mexico

A total of 96 epiphytic algae species were identified from Bajo Pepito, Quintana Roo, México. 60.4% (58) belonged to the Rhodophyta, 19.79% (19) to the Phaeophyta, 16.6% (16) to the Chlorophyta and 3.1% (3) to the Cyanophyta; 49 species (50.5%) were found only in one month, while Heterosiphonia crispella was found in all of the sampled months. That species provided the largest contribution to the biomass of epiphytes. During January we registered the greater biommass and richness of epiphytes species, coincidently with high values of host species cover and rainfall.     

C-phycocyanin and allophycocyanin in two species of blue-green algae

1. The biliproteins C-phycocyanin and allophycocyanin were purified from the blue-green alga Anabaena variabilis by ammonium sulphate fractionation and gel filtration. 2. An assay procedure that enabled the proportion of the two pigments, present as a mixture, to be determined was devised by using the data provided by spectrophotometric analysis of the purified biliproteins. 3. The degree of association and relative proportions of the two pigments were analysed by the application of this procedure to the components separated by thin-layer gel filtration. 4. The C-phycocyanin/allophycocyanin ratio remained essentially constant in algal extracts prepared at various stages throughout the growth cycle or after growth under conditions of reduced illumination. 5. The behaviour of the C-phycocyanin aggregate species from Anacystis nidulans suggested that they were of appreciably lower molecular weight than those observed in extracts of Anabaena variabilis.     

Assessment of the seaweed-seagrass resource of Mararison Island,Culasi, Antique,Philippines*

A bimonthly sampling of the seaweed-seagrass resource of Mararison Island, Culasi Antique, was undertaken over 1 year to assess the species composition, similarity of taxa, and biomass (dry weight [d.w.] g m^?2) at seven localities. A total of 45 species was identified: 17 Chlorophyta, seven Phaeophyta, 15 Rhodophyta, one Cyanophyta and five seagrasses. Except for some Rhodophyta and Syringodium isoetifolium (Ascherson) Dandy, the occurrence of species between stations was not significantly different; however, differences in biomass between sampling time (month) were significant. Identical taxa between stations were determined. The highest (40) and lowest (22) number of species collected were in May and July, respectively. The species were most abundant from March to May (dry months) and sparse from July to September (wet months). The most abundant species were: Sargassum polycystum C. Agardh (399 g m^?2) (Phaeophyta), Dictyosphaeria cav-ernosa (Forsskat) Borgesen (43.1 g m^?2) (Chlorophyta), Acanthopeitis japonica Okamura (97.2 gm^?2) (Rhodophyta) and Thalassia hemprichii (Ehrenberg) Ascherson (1370 g m^?2; seagrass). The Phaeophyta were abundant in March, and the Chlorophyta and Rhodophyta in May, while the seagrasses were abundant in September. Some species occurred only during the dry months: two Phaeophyta, nine Chlorophyta and five Rhodophyta. All the seagrasses were found year-round. Almost all of the seaweeds (39/45) were found associated with seagrass. The number of seaweeds in Mararison Island was higher than for seagrasses but the total biomass of the latter was much higher than the combined biomass of the seaweeds.     

New records of Malaysian marine algae

Checklists of the marine algae of Malaysia have been published. The last checklist included the marine algae of Singapore. The checklist recorded 212 taxa for the region. This paper lists an additional 7 taxa of Chlorophyta, 18 taxa of Rhodophyta and 5 taxa of Phaeophyta, which had been left out of the last checklist. These include marine algae from Pulau Pinang, Pulau Redang and Pulau Sibu. In addition, new records of Cyanophyta (2 species), Chlorophyta (4 species), Rhodophyta (10 species) and Phaeophyta (3 species) are reported.     

福建崇武沿海野生经济底栖海藻资源

福建崇武沿海有底栖海藻46种。其中野生经济底栖海藻34种(蓝藻门1种,绿藻门8种,红藻门17种,褐藻门8种)。文中对这些海藻的经济价值及资源开发利用进行阐述。     

MOLECULAR PHYLOGENY OF PHYCOCYANIN‐CONTAINING CRYPTOPHYTES: EVOLUTION OF BILIPROTEINS AND GEOGRAPHICAL DISTRIBUTION1

Of 34 strains assigned to the cryptophyte genera Chroomonas Hansg., Hemiselmis Parke, and Komma D. R. A. Hill, distribution patterns of biliproteins, habitats, and sampling sites across a phylogenetic tree have been examined. The combined data set assembled from nuclear SSU rDNA, partial nuclear LSU rDNA, and nucleomorph SSU rDNA sequences comprised 4,083 positions and yielded an almost completely resolved tree. Spectrophotometry of the biliproteins and mapping of the different types of biliproteins onto the phylogenetic tree unveiled a complex evolutionary history. Different from other cryptophyte clades, the types of biliproteins were not generally congruent with clades or subclades of the genera Chroomonas (paraphyletic, phycocyanins [PCs] 645 or 630), Hemiselmis (PCs 612, 630 or phycoerythrin [PE] 555), and Komma (PC 645). At least one putative character reversal took place in the genus Chroomonas. Several changes in biliproteins have been found in the genus Hemiselmis, including two new biliprotein variants that probably originated by slight modifications from PC 612 and PE 555, respectively (PC 577 and PE 545/555). Freshwater and marine/brackish taxa were intermingled across the tree without displaying a specific pattern. In four terminal clades, genetically identical strains have been found to occur both in Europe and in the USA. The Chroomonas/Hemiselmis/Komma clade proved to be the most diverse of all cryptophyte clades concerning types of biliproteins and distribution of clades across marine or freshwater habitats.     

Characterization of phycoerythrin from a Cryptomonas sp.

Summary Two closely similar phycoerythrins were purified from Cryptomonas sp. The two proteins were indistinguishable with respect to native molecular weight, subunit structure, photolability and immunological specificity, and differed only in their isoelectric points (pH 5.74 and 6.35), as determined by isoelectric focussing in polyacrylamide gels. Each protein consisted of two unequal subunits, (mol. wt. 11,800) and (mol. wt. 19,000), and each subunit contained covalently bound chromophore. In contrast to the blue-green and red algal phycoerythrins studied thus far, the Cryptomonas sp. phycoerythrins are extremely photolabile; exposure of the purified proteins to relatively short periods of intense illumination with visible light produces a marked decrease in fluorescence and in absorbance at 567 m.Abbreviation used SDS sodium dodecyl sulfate     

Choline and inositol distribution in algae and fungi

Inositol and choline were present in varying amounts among the species of Rhodophyta, Phaeophyta, Chlorophyta, and Euglenophyta examined. However, in the two members of the order Fucales (division Phaeophyta) examined, no detectable amounts of choline were found. In contrast, the species of Cyanophyta examined contained no detectable amounts of either choline or inositol. All species of the fungal classes Phycomyceteae, Ascomyceteae, and Basidiomyceteae collected contained both inositol and choline in varying amounts. The red, brown, and blue-green algae usually contained much less inositol and choline than do plant and animals sources, but the fungi and the algae Chlorella and Euglena contained amounts comparable to those present in plant sources.