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D A Bryant

Publications and source records attributed to D A Bryant.

83 records · Page 5Linked to original sources
PubMed1985

Molecular cloning and nucleotide sequence of the alpha and beta subunits of allophycocyanin from the cyanelle genome of Cyanophora paradoxa.

The genes for the alpha- and beta-subunit apoproteins of allophycocyanin (AP) were isolated from the cyanelle genome of Cyanophora paradoxa and subjected to nucleotide sequence analysis. The AP beta-subunit apoprotein gene was localized to a 7.8-kilobase-pair Pst I restriction fragment from cyanelle DNA by hybridization with a tetradecameric oligonucleotide probe. Sequence analysis using that oligonucleotide and its complement as primers for the dideoxy chain-termination sequencing method confirmed the presence of both AP alpha- and beta-subunit genes on this restriction fragment. Additional oligonucleotide primers were synthesized as sequencing progressed and were used to determine rapidly the nucleotide sequence of a 1336-base-pair region of this cloned fragment. This strategy allowed the sequencing to be completed without a detailed restriction map and without extensive and time-consuming subcloning. The sequenced region contains two open reading frames whose deduced amino acid sequences are 81-85% homologous to cyanobacterial and red algal AP subunits whose amino acid sequences have been determined. The two open reading frames are in the same orientation and are separated by 39 base pairs. AP alpha is 5' to AP beta and both coding sequences are preceded by a polypurine, Shine-Dalgarno-type sequence. Sequences upstream from AP alpha closely resemble the Escherichia coli consensus promoter sequences and also show considerable homology to promoter sequences for several chloroplast-encoded psbA genes. A 56-base-pair palindromic sequence downstream from the AP beta gene could play a role in the termination of transcription or translation. The allophycocyanin apoprotein subunit genes are located on the large single-copy region of the cyanelle genome.

Amino Acid Sequence↗
PubMed1985

Gene map for the Cyanophora paradoxa cyanelle genome.

The genes for the following proteins were localized by hybridization analysis on the cyanelle genome of Cyanophora paradoxa: the alpha and beta subunits of phycocyanin (cpcA and cpcB); the alpha and beta subunits of allophycocyanin (apcA and apcB); the large and small subunits of ribulose-1,5-bisphosphate carboxylase (rbcL and rbcS); the two putative chlorophyll alpha-binding apoproteins of the photosystem I-P700 complex (psaA and psaB); four apoproteins believed to be components of the photosystem II core complex (psbA, psbB, psbC, and psbD); the two apoprotein subunits of cytochrome b-559 which is also found in the core complex of photosystem II (psbE and psbF); three subunits of the ATP synthase complex (atpA and atpBE); and the cytochrome f apoprotein (petA). Eighty-five percent of the genome was cloned as BamHI, BglII, or PstI fragments. These cloned fragments were used to construct a physical map of the cyanelle genome and to localize more precisely some of the genes listed above. The genes for phycocyanin and allophycocyanin were not clustered and were separated by about 25 kilobases. Although the rbcL gene was adjacent to the atpBE genes and the psbC and psbD genes were adjacent, the arrangement of other genes encoding various polypeptide subunits of protein complexes involved in photosynthetic functions was dissimilar to that observed for known chloroplast genomes. These results are consistent with the independent development of this cyanelle from a cyanobacterial endosymbiont.

Animals↗
PubMed1984

Genes for the alpha and beta subunits of phycocyanin.

Phycocyanin (PC) is a light-harvesting protein common to blue-green and red algae. We have isolated the genes for the two apoprotein subunits, alpha and beta, of PC from the blue-green alga Agmenellum quadruplicatum PR-6. We synthesized eight sense-strand tetradecameric oligonucleotide probes that could encode a particular pentapeptide in PC alpha from A. quadruplicatum. Only one probe hybridized with total RNA from this organism. This oligonucleotide showed homology to a unique restriction fragment when used to probe Southern blots of A. quadruplicatum DNA. The probe-homologous 3.1-kilobase pair HindIII fragment was cloned. The nucleotide sequence of a 1.7-kilobase pair segment of this clone was determined. Two open reading frames are contained in this region, which correspond in deduced amino acid sequence to PC alpha and PC beta subunits. Both coding sequences are in the same orientation, separated by 105 base pairs, with the PC beta gene 5' to the PC alpha gene. Each gene has a Shine-Dalgarno-type sequence near the initiation codon. Codon frequencies in the two genes may be correlated with the abundance of their products. The deduced amino acid sequences of the gene products show considerable homology with the alpha and beta PC subunits from other species.

Amino Acid Sequence↗
PubMed1982

Growth and Chromatic Adaptation of Nostoc sp. Strain MAC and the Pigment Mutant R-MAC.

A spontaneous, stable, pigmentation mutant of Nostoc sp. strain MAC was isolated. Under various growth conditions, this mutant, R-MAC, had similar phycoerythrin contents (relative to allophycocyanin) but significantly lower phycocyanin contents (relative to allophycocyanin) than the parent strain. In saturating white light, the mutant grew more slowly than the parent strain. In nonsaturating red light, MAC grew with a shorter generation time than the mutant; however, R-MAC grew more quickly in nonsaturating green light.When the parental and mutant strains were grown in green light, the phycoerythrin contents, relative to allophycocyanin, were significantly higher than the phycoerythrin contents of cells grown in red light. For both strains, the relative phycocyanin contents were only slightly higher for cells grown in red light than for cells grown in green light. These changes characterize both MAC and R-MAC as belonging to chromatic adaptation group II: phycoerythrin synthesis alone photocontrolled.A comparative analysis of the phycobilisomes, isolated from cultures of MAC and R-MAC grown in both red and green light, was performed by polyacrylamide gel electrophoresis in the presence of 8.0 molar urea or sodium dodecyl sulfate. Consistent with the assignment of MAC and R-MAC to chromatic adaptation group II, no evidence for the synthesis of red light-inducible phycocyanin subunits was found in either strain. Phycobilisomes isolated from MAC and R-MAC contained linker polypeptides with relative molecular masses of 95, 34.5, 34, 32, and 29 kilodaltons. When grown in red light, phycobilisomes of the mutant R-MAC appeared to contain a slightly higher amount of the 32-kilodalton linker polypeptide than did the phycobilisomes isolated from the parental strain under the same conditions. The 34.5-kilodalton linker polypeptide was totally absent from phycobilisomes isolated from cells of either MAC or R-MAC grown in green light.

Journal Article↗
PubMed1981

Differential thermal analysis of dipalmitoylphosphatidylcholine--fatty acid mixtures.

Mixtures of dipalmitoylphosphatidylcholine (DPPC) with palmitic, stearic, and myristic acids and the sodium salts of these acids were analyzed by differential thermal analysis (DTA) over a wide range of lipid compositions, all in excess water. All three fatty acids raise the liquid-crystal phase transition temperature and form sharp-melting complexes, with 1:2 DPPC--fatty acid stoichiometry observed for palmitic and stearic acids and suggested for myristic acid. Phase diagrams of the peritectic type, indicating nonideal mixing, was fitted to the DPPC--palmitic acid and DPPC--stearic acid data. In contrast, DPPC forms nearly ideal mixtures with the putative DPPC--myristic acid complex. At levels of only a few mole percent, both sodium stearate and myristate remove the pretransition and main transition and produce new peaks at approximately 30 and approximately 48 degrees C; the relative areas of the new peaks were unreproducible for the DPPC--myristate system. Sodium palmitate is the least disruptive of any of the sodium soaps or fatty acids; up to 80 mol % palmitate, the transition is lowered 3 degrees C and approximately doubled in width. The pretransition is detectable up to 36 mol %, and the main transition persists up to 88 mol % palmitate. The apparent pK of palmitic acid (12 mol %) in DPPC bilayers was determined to be 10.2 by direct pH measurement of ternary DPPC mixtures with known palmitic acid/sodium palmitate ratios; the intrinsic pK is estimated to be less than or approximately 8.5.

Fatty Acids, Nonesterified↗
PubMed1981

Effects of chromatic illumination on cyanobacterial phycobilisomes. Evidence for the specific induction of a second pair of phycocyanin subunits in Pseudanabaena 7409 grown in red light.

Pseudanabaena 7409 is a chromatically cyanobacterium which photocontrols the synthesis of both phycoerythrin and phycocyanin [Tandeau de Marsac (1977) J. Bacteriol. 130, 82--91]. Phycobilisomes, isolated from cells grown in either green or red light, have been dissociated and the component biliproteins purified and characterized. Phycobilisomes isolated from cells grown in green light were composed of allophycocyanin B, allophycocyanin, two phycocyanin subunits (one alpha-type and one beta-type subunit), phycoerythrin and eight uncolored polypeptides. When dissociated phycobilisomes were chromatographed on DEAE-cellulose at pH 5.5, most of the phycocyanin was recovered as part of a large (17.3 S) multiprotein complex with phycoerythrin (molar ratio 1 : 1). This complex also contained five of the uncolored polypeptides found in intact phycobilisomes isolated from cells grown in green light. Phycobilisomes isolated from cells grown in red light were composed of allophycocyanin B, allophycocyanin, four phycocyanin subunits (two alpha-type and two beta-type subunits), and six uncolored polypeptides. When these phycobilisomes were dissociated, the phycocyanin was recovered as a large (21.0 S) multiprotein complex which was composed of the four phycocyanin subunits types and four uncolored polypeptides. This complex was morphologically identical to the rod-like stacks of discs about 6 x 12 nm which form the peripheral rods of intact phycobilisomes. Each of the four phycocyanin subunits found in the complex isolated from the phycobilisomes of cells grown in red light was purified to homogeneity and characterized. Amino acid compositions of the four subunits indicated that each subunit was a unique gene product. Two of the subunits of the complex were apparently identical to those of the phycocyanin purified from phycobilisomes isolated from cells grown in green light. These studies suggest that one pair of phycocyanin subunits was synthesized constitutively (i.e. irrespective of the light wavelength to which the cells were exposed during growth) while the synthesis of the second pair of phycocyanin subunits was specifically induced during growth in red light.

Amino Acids↗
PubMed1981

The photoregulated expression of multiple phycocyanin species. A general mechanism for the control of phycocyanin synthesis in chromatically adapting cyanobacteria.

The regulation of phycocyanin synthesis in response to growth in chromatic illumination was studied in 69 strains of cyanobacteria. Cyanobacteria (24 of 31 strains examined), which chromatically adapt by modulating the synthesis of both phycocyanin and phycoerythrin, controlled phycocyanin synthesis through the differential, photoregulated expression of two phycocyanin species (two alpha-type and two beta-type subunits). For these strains the expression of one pair of phycocyanin subunits was constitutive (i.e. irrespective of the light wavelength in which the cells were grown); the expression of the second pair of phycocyanin subunits occurred specifically during growth in red light. Two facultatively heterotrophic cyanobacteria, Calothrix strains 7101 and 7601, synthesized both the constitutive and the inducible pairs of phycocyanin subunits when grown heterotrophically in the dark after transfer from either red or green light. No evidence for the existence of multiple and/or photoregulated phycocyanin species was found for cyanobacteria (25 strains) incapable of chromatic adaptation, nor for cyanobacteria (13 strains) which chromatically adapt by modulating the synthesis of phycoerythrin alone.

Acclimatization↗
PubMed1976

Characterization and structural properties of the major biliproteins of Anabaena sp.

Studies are presented of the biliproteins of Anabaena sp. This filamentous cyanobacterium contains three major biliproteins. Whereas two of these, C-phycocyanin and allophycocyanin, are common to all cyanobacteria, the third, phycoerythrocyanin (gammamax approximately 568 nm) has hitherto not been described and its distribution among cyanobacteria appears to be limited. Anabaena variabilis and Anabaena sp. 6411 allophycocyanin, C-phycocyanin, and phycoerythrocyanin were purified to homogeneity and characterized with respect to molecular weight, isoelectric point, absorption spectrum and amino acid composition. The alpha and beta subunits of each of these proteins were also purified to homogeneity and characterized in the same manner. The tetrapyrrole chromophore content was determined for each of the proteins and subunits. The alpha subunit of phycoerythrocyanin carries a novel phycobiliviolin-like chromophore. This chromophore has not previously been detected in cyanobacterial biliproteins, but has been noted as a prosthetic group of a cryptophytan phycocyanin. Sedimentation equilibrium studies show that at pH 7.0, at protein concentrations of 0.2-0.6 mg/ml, allophycocyanin, C-phycocyanin and phycoerythrocyanin, each exists as a trimeric aggregate, (alphabeta)3, of molecular weight of approximately 105000. Structrual studies of microcrystals of these three biliproteins by electron microscopy and X-ray diffraction reveal a common plan for the construction of higher assembly forms. The major building block appears to be the trimer (alphabeta)3. It is proposed that this is a disc-like structure about 3.0 X 12.0 nm. The individual alpha or beta subunits are roughly spherical, 3 nm in diameter. Allophycocyanin trimers stack to form bundles of rods which form long needles. Both phycocyanin and phycoerythrocyanin form double discs (alphabeta)6 which are visible as ring-shaped structures by electron microscopy. The mode of assembly of the biliprotein structures in the phycobilisome is, as yet, unknown.

Amino Acids↗
PubMed1976

Biliproteins of cyanobacteria and Rhodophyta: Homologous family of photosynthetic accessory pigments.

Amino-terminal sequence determinations are reported of the subunits of biliproteins of prokaryotic unicellular and filamentous cyanobacteria and of eukaryotic unicellular red algae. The biliproteins examined, allophycocyanin, C-phycocyanin, R-phycocyanin, b-phycoerythrin, and phycoerythrocyanin, vary with respect to the chemical nature and the number and distribution of the bilin chromophores between the two dissimilar subunits. The amino-terminal sequences fall into two classes, "alpha-type" and "beta-type", with a high degree of homology within each class.In those biliproteins where the number of bilin chromophores on the two subunits is unequal, the subunit with the greater number of chromophores has the beta-type amino-acid sequence.Extensive homology also exists between alpha- and beta-type sequences, strongly supporting the view that these arose by gene duplication to give rise to the ancestral alpha- and beta-type genes early in the evolution of the biliproteins. The subsequent generation of the various classes of biliproteins appears to be the result of further gene duplication of the alpha- and beta-type genes, ultimately to give rise to families of polypeptide chains of similar sequence, but varying in the number of chromophore attachment sites and the structure of the chromophores.

Journal Article↗
PubMed1975

Allophycocyanin B (lambdamax 671, 618 nm): a new cyanobacterial phycobiliprotein.

A hitherto undescribed red fluorescent phycobiliprotein (maximum emission at congruent to 680 nm), characterized by long wavelength absorption maxima in the visible region at 671 nm (xi = 172000 M(-1).cm(-1) per monomer of mol. wt. 30600)and 618 nm, has been purified to homogeneity from unicellular cyanobacterium, Synechococcus sp., and from a filamentous cyanobacterium, Anabaena variabilis. The name allophycocyanin B has been proposed for the new protein. A. variabilis allophycocyanin B is characterized by a native molecular weight of 89000 p 5000 (in 0.05 M phosphate at pH 7.2), an isoelectric point of 5.09, and a subunit molecular weight, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, of 15300. The protein contains one phycocyanobilin chromophore per subunit. In common with allophycocyanin from the same organism, allophycocyanin B does not contain either histidine or tryptophan. In other respects, the amino acid compositions of the two proteins are significantly different. Synechococcus sp. (Anacytis nidulans) allophycocyanin B gives two components of 16000 and 17000 mol. wt., of equal staining intensity, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Allophycocyanin B from both organisms cross-react with rabbit antisera directed against either Synechococcus sp. or Anabaena sp. allophycocyanin, but not with antisera against the phycocyanins of the same organisms. It is suggested that allophycocyanin B occupoes a position between allophycocyanin and chlorophyll a in the energy transfer path from the accessory pigments to species of chlorophyll a with absorption maxima at lambda greater than 670 nm.

Amino Acids↗