Isolation, characterization and electron microscopy analysis of a hemidiscoidal phycobilisome type from the cyanobacterium Anabaena sp. PCC 7120.

In this work we present the characterization of a hemidiscoidal phycobilisome type of the heterocyst-forming cyanobacterium Anabaena sp. PCC 7120. The phycobilisome of this organism contains allophycocyanin, phycocyanin and phycoerythrocyanin, similar to the closely related thermophilic cyanobacterium Mastigocladus laminosus. Intact phycobilisomes exhibit an absorption maximum at 619 nm and two fluorescence maxima at 664 nm and 680 nm, corroborating the presence of a complete energy pathyway along the antenna. Upon dissociation, the phycobiliproteins were released from the phycobilisome. One phycoerythrocyanin, one phycocyanin and three allophycocyanin complexes were isolated by ion-exchange chromatography and characterized by absorption and fluorescence spectroscopy and by SDS/PAGE. The amino-terminal sequences of the polypeptides belonging to the phycoerythrocyanin and phycocyanin families were identical with the derived sequences of their corresponding genes. Partial amino-terminal sequences of the polypeptides belonging to the allophycocyanin family are presented here. Our results show that the phycobiliproteins and linker polypeptides from Anabaena sp. PCC 7120 are similar to the phycobilisome components characterized in other cyanobacteria. The phycobilisome of Anabaena sp. PCC 7120 was extensively analyzed by electron microscopy. It differs from the common hemidiscoidal tricylindrical, six-rod phycobilisome type by a core domain consisting of five core cylinders surrounded by up to eight rods radiating in a hemidiscoidal manner. One rod is linked to each basal core cylinder, whereas the remaining core cylinders bind two rods each. On the basis of the data presented in this work, a revised model for the hemidiscoidal pentacylindrical phycobilisome of Anabaena sp. PCC 7120, M. laminosus and Anabaena variabilis is proposed. This model accounts more accurately for the 'grape' pattern typically exhibited by these phycobilisomes in electron micrographs.

[1]  H. Zuber,et al.  Structure of light-harvesting antenna complexes of photosynthetic bacteria, cyanobacteria and red algae , 1986 .

[2]  B. Zilinskas,et al.  Allophycocyanin from Nostoc sp. phycobilisomes. Properties and amino acid sequence at the NH2 terminus of the alpha and beta subunits of allophycocyanins I, II, and III. , 1980, The Journal of biological chemistry.

[3]  R. Huber,et al.  Isolation, crystallization, crystal structure analysis and refinement of constitutive C-phycocyanin from the chromatically adapting cyanobacterium Fremyella diplosiphon at 1.66 A resolution. , 1991, Journal of molecular biology.

[4]  R. Huber,et al.  Refined three-dimensional structures of two cyanobacterial C-phycocyanins at 2.1 and 2.5 A resolution. A common principle of phycobilin-protein interaction. , 1987, Journal of molecular biology.

[5]  H. Zuber,et al.  The complete amino acid sequence of R‐phycocyanin‐I α and β subunits from the red alga Porphyridium cruentum , 1994 .

[6]  A. Glazer,et al.  Allophycocyanin B. A common beta subunit in Synechococcus allophycocyanin B (lambda max 670 nm) and allophycocyanin (lambda max 650 nM). , 1981, The Journal of biological chemistry.

[7]  V. Capuano,et al.  The "anchor polypeptide" of cyanobacterial phycobilisomes. Molecular characterization of the Synechococcus sp. PCC 6301 apce gene. , 1991, The Journal of biological chemistry.

[8]  A. Glazer,et al.  Molecular architecture of a light-harvesting antenna. Structure of the 18 S core-rod subassembly of the Synechococcus 6301 phycobilisome. , 1983, The Journal of biological chemistry.

[9]  D. Bryant CHAPTER 7 – Cyanobacterial Phycobilisomes: Progress toward Complete Structural and Functional Analysis via Molecular Genetics , 1991 .

[10]  A. Glazer Phycobilisome a macromolecular complex optimized for light energy transfer , 1984 .

[11]  H. Zuber,et al.  Chapter 11 Structure and function of light-harvesting pigment-protein complexes , 1987 .

[12]  H. Zuber,et al.  Linker polypeptides of the phycobilisome from the cyanobacterium Mastigocladus laminosus. I. Isolation and characterization of phycobiliprotein-linker-polypeptide complexes. , 1986, Biological chemistry Hoppe-Seyler.

[13]  A. Glazer,et al.  Molecular architecture of a light-harvesting antenna. Comparison of wild type and mutant Synechococcus 6301 phycobilisomes. , 1980, The Journal of biological chemistry.

[14]  H. Zuber,et al.  The phycobiliprotein beta 16.2 of the allophycocyanin core from the cyanobacterium Mastigocladus laminosus. Characterization and complete amino-acid sequence. , 1987, Biological chemistry Hoppe-Seyler.

[15]  H. Zuber,et al.  Phycobilisome structure in the cyanobacteria Mastigocladus laminosus and Anabaena sp. PCC 7120. , 1992, European journal of biochemistry.

[16]  R. Huber,et al.  Crystal structure analysis and refinement at 2.5 A of hexameric C-phycocyanin from the cyanobacterium Agmenellum quadruplicatum. The molecular model and its implications for light-harvesting. , 1986, Journal of molecular biology.

[17]  W. Sidler,et al.  Phycobilisome and Phycobiliprotein Structures , 1994 .

[18]  H. Zuber,et al.  A small multigene family encodes the rod-core linker polypeptides of Anabaena sp. PCC7120 phycobilisomes. , 1991, Gene.

[19]  F. Lottspeich,et al.  ORIGIN OF THE RED SHIFTED ABSORPTION IN PHYCOCYANIN 632 FROM Mastigocladus laminosus , 1991 .

[20]  R. Huber,et al.  X-ray crystallographic structure of the light-harvesting biliprotein C-phycocyanin from the thermophilic cyanobacterium Mastigocladus laminosus and its resemblance to globin structures. , 1985, Journal of molecular biology.

[21]  D. Bryant,et al.  Molecular characterization of ferredoxin-NADP+ oxidoreductase in cyanobacteria: cloning and sequence of the petH gene of Synechococcus sp. PCC 7002 and studies on the gene product. , 1992, Biochemistry.

[22]  A. Glazer,et al.  Genes encoding the phycobilisome rod substructure are clustered on the Anabaena chromosome: characterization of the phycoerythrocyanin operon , 1992, Journal of bacteriology.

[23]  A. Glazer,et al.  Light guides. Directional energy transfer in a photosynthetic antenna. , 1989, The Journal of biological chemistry.

[24]  H. Zuber,et al.  Structure of the genes encoding the rod-core linker polypeptides of Mastigocladus laminosus phycobilisomes and functional aspects of the phycobiliprotein/linker-polypeptide interactions. , 1992, European journal of biochemistry.

[25]  M. Mimuro,et al.  Allophycocyanin complexes of the phycobilisome from Mastigocladus laminosus. Influence of the linker polypeptide L8.9C on the spectral properties of the phycobiliprotein subunits. , 1987, Biological chemistry Hoppe-Seyler.

[26]  Herbert Zuber,et al.  STRUCTURE AND FUNCTION OF LIGHT‐HARVESTING COMPLEXES AND THEIR POLYPEPTIDES , 1985 .

[27]  R. Huber,et al.  Isolation, crystallization, crystal structure analysis and refinement of B-phycoerythrin from the red alga Porphyridium sordidum at 2.2 A resolution. , 1992, Journal of molecular biology.

[28]  D. Bryant Phycoerythrocyanin and Phycoerythrin: Properties and Occurrence in Cyanobacteria , 1982 .

[29]  H. Zuber,et al.  ISOLATION, CHARACTERIZATION AND RECONSTITUTION OF PHYCOBILIPROTEIN ROD‐CORE LINKER POLYPEPTIDE COMPLEXES FROM THE PHYCOBILISOME OF Mastigocladus laminosus * , 1993 .

[30]  T. Katoh,et al.  Subparticles of Anabaena Phycobilisomes I. : Reconstitution of Allophycocyanin Cores and Entire Phycobilisomes , 1983 .

[31]  V. Capuano,et al.  Isolation and molecular characterization of the gene encoding allophycocyanin B, a terminal energy acceptor in cyanobacterial phycobillsomes , 1988, Molecular microbiology.

[32]  H. Zuber,et al.  Three C‐phycoerythrin‐associated linker polypeptides in the phycobilisome of green‐light‐grown Calothrix sp. PCC 7601 (cyanobacteria) , 1992, FEBS letters.

[33]  B. Zilinskas,et al.  Role of the colorless polypeptides in phycobilisome reconstitution from separated phycobiliproteins. , 1982, Plant physiology.

[34]  A. Glazer,et al.  Molecular architecture of a light-harvesting antenna. Core substructure in Synechococcus 6301 phycobilisomes: two new allophycocyanin and allophycocyanin B complexes. , 1983, The Journal of biological chemistry.

[35]  V. Capuano,et al.  Molecular characterization of the terminal energy acceptor of cyanobacterial phycobilisomes. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[36]  A. Grossman Chromatic adaptation and the events involved in phycobilisome biosynthesis , 1990 .

[37]  E. Gantt,et al.  Phycobilisomes from blue-green and red algae: isolation criteria and dissociation characteristics. , 1979, Plant physiology.

[38]  H. Siegelman,et al.  Preparation of calcium phosphate for protein chromatography. , 1965, Analytical biochemistry.

[39]  R. Huber,et al.  Refined three-dimensional structure of phycoerythrocyanin from the cyanobacterium Mastigocladus laminosus at 2.7 A. , 1990, Journal of molecular biology.

[40]  T. Katoh,et al.  Subparticles of Anabaena phycobilisomes. II. Molecular assembly of allophycocyanin cores in reference to "anchor" protein. , 1987, Archives of biochemistry and biophysics.

[41]  D. Bryant,et al.  Genetic analysis of a 9 kDa phycocyanin-associated linker polypeptide. , 1990, Biochimica et biophysica acta.

[42]  W. Wehrmeyer Phycobilisomes: Structure and function , 1990 .

[43]  M. Mimuro,et al.  Energy flow in the phycobilisome core of Nostoc sp. (MAC): Two independent terminal pigments , 1986 .

[44]  A. Glazer,et al.  Molecular architecture of a light-harvesting antenna. Quaternary interactions in the Synechococcus 6301 phycobilisome core as revealed by partial tryptic digestion and circular dichroism studies. , 1983, The Journal of biological chemistry.

[45]  A. Glazer,et al.  Cyanobacterial Phycobilisomes. Particles from synechocystis 6701 and two pigment mutants , 1980, The Journal of cell biology.

[46]  A. Glazer,et al.  Core substructure in cyanobacterial phycobilisomes , 1983, Journal of cellular biochemistry.

[47]  A. Glazer,et al.  Structural studies on phycobiliproteins III. Comparison of bilin-containing peptides from the beta subunits of C-phycocyanin, R-phycocyanin, and phycoerythrocyanin. , 1978, The Journal of biological chemistry.

[48]  R. Haselkorn,et al.  Cloning and light regulation of expression of the phycocyanin operon of the cyanobacterium Anabaena. , 1987, The EMBO journal.

[49]  H. Zuber,et al.  Minor Polypeptides from the Phycobilisome of the Cyanobacterium Mastigocladus laminosus. Isolation, Characterization and Amino-Acid Sequences of a Colourless 8.9-kDa Polypeptide and of a 16.2-kDa Phycobiliprotein , 1984 .

[50]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.