Diversity in the monosaccharide composition of antigenic polysaccharides from proteobacteria Pseudoalteromonas and Marinomonas genera]

The sugar analysis of the glycans of the type strains of marine proteobacteria of the genera Pseudoalteromonas and Marinomonas--Pseudoalteromonas atlantica IAM12927T, P. aurantia NCIMB 2033T, P. citrea ATCC 29719T, P. elyakovii KMM 162T, P. espejiana ATCC 29659T, P. piscicida NCIMB 645T, P. tetraodonis IAM 14160T, Marinomonas communis ATCC 27118T, and M. vaga ATCC 27119T--showed that they contain glucose, galactose, galactosamine, glucosamine, fucose, rhamnose, mannose, heptose, 2-keto-3-deoxyoctonate (KDO), uronic acids, colitose (3,6-dideoxyl-L-xylo-hexose), and 6-deoxy-L-talose. The carbohydrate composition of the antigenic polysaccharides (PSs) of P. elyakovii KMM 162T and P. espejiana ATCC 29659T depended on the type and the concentration of carbohydrate substrates in the nutrient media. The molar proportion between rhamnose, glucose, and galactose (ca. 1:0.3:2) in the PS of P. elyakovii KMM 162T was almost the same in the media lacking carbohydrates or containing glucose or galactose at a concentration of 1 g/l. At the same time, the molar proportion between fucose, glucose, galactose, galactosamine, and glucosamine (ca. 1:1:1:2:0.5) in the PS of P. espejiana ATCC 29659T depended on the presence and the concentration of carbohydrate substrates in the medium. A high concentration of glucose in the medium (30 g/l) brought about a rise in the content of glucose in PSs (9-fold for the PS of P. elyakovii KMM 162T and 4.6-fold for the PS of P. espejiana ATCC 29659T) and led to a decrease in the content of other carbohydrates. The cultivation of these two strains at a lactose concentration of 30 g/l resulted in their PSs containing glucose and galactose in about equal proportions (ca. 1:1 in the case of P. espejiana ATCC 29659T and ca. 2.1:1.7 in the case of P. elyakovii KMM 162T).     

Different polysaccharides in the external layers (capsule and slime) of the cell envelope of Rhodopseudomonas capsulata Sp11

Two different acidic polysaccharides (I and II) were detected in the external cell envelope layers (slime and capsule) of Rhodopseudomonas capsulata Sp11. Polysaccharide I contains rhamnose, fucose, glucosamine and an unknown acidic sugar, it represents the slime material of the strain. Polysaccharide II contains rhamnose, galactose, 3-amino-3,6-dideoxygalactose, an unknown amino sugar and galacturonic acid, it represents very likely the capsule of R. capsulata Sp11. Polysaccharide I has a serological specificity different from that of polysaccharide II as shown by immunoprecipitation using antisera against living cells. Polysaccharide II, but not polysaccharide I, reacts in antiserum against heat-treated cells (100 degrees C, 2.5 h). Whole cells are agglutinated in the antisera against living but not in those against heat-treated cells.     

Trypsin Inhibitor Polymorphism: Multigene Family Expression and Posttranslational Modification

Trypsin inhibitors from winter pea seeds (c.v. Frilene) have been purified and shown to consist of six protease inhibitors (PSTI I, II, III, IVa, IVb, and V). Based on amino acid composition, molecular mass, and N-terminal sequence, the six inhibitors are closely related to one another and belong to the Bowman–Birk family of inhibitors. To define the relations among them, molecular mass and amino acid composition of peptides obtained from digestion with trypsin were determined. The sequence and the biosynthetic mechanism of the isoform formation have been partially resolved for four major isoforms. Two isoinhibitor forms (PSTI IVa, IVb) in pea seeds are due to expression of two distinct genes; PSTI IVa has four amino acid replacements when its sequence is compared with the sequence of PSTI IVb. Two others (PSTI I, II) result from posttranslational proteolytic cleavage of nine C-terminal residues of forms PSTI IVa and IVb, respectively.     

Structures of oligosaccharides on beta-galactosidase from Aspergillus oryzae

The carbohydrate portions of beta-galactosidase from Aspergillus oryzae were found to be composed of two types of sugar chains. They were released equally well with endo-beta-N-acetylglucosaminidase H, but were distinct in their chain length. The long sugar chains (fraction I), corresponding to 4% of the total carbohydrate chains, were composed of galactomannan-type oligosaccharides, which consisted of mannose, galactose, glucose, and glucosamine in the molar ratios of 30.0, 16.4, 1.4, and 2.1 per mol of aspartic acid, respectively. The short sugar chains (fraction II), corresponding to 96% of the total carbohydrate chains, consisted of mannose, galactose, glucose, and glucosamine in the molar ratios of 9.4, 0.6, 0.3, and 1.7 per mol of aspartic acid, respectively. Both types of sugar chains were fractionated into neutral and acidic subfractions. The neutral subfraction of fraction I (I-N), corresponding to 1% of the total carbohydrate chains, was very heterogeneous in length and was resistant to digestion with alpha-mannosidase and beta-galactosidase. The neutral subfraction of fraction II (II-N), corresponding to 91% of the total carbohydrate, was composed of a mixture of oligosaccharides with oligomanneoside chains (Mann GlcNAcol). The major components were similar to high mannose-type oligosaccharides of mammalian origin in their composition and size (n = 5-9). However, digestion of II-N with alpha 1,2-mannosidase produced considerable amounts of Man6GlcNAcol, an unusual product in the case of high mannose-type oligosaccharides of mammalian origin, in addition to the common one, Man5GlcNAcol.     

Diversity of the Carbohydrate Composition of the Antigenic Polysaccharides of Proteobacteria of the Genera Pseudoalteromonasand Marinomonas

The sugar analysis of the glycans of the type strains of marine proteobacteria of the genera Pseudoalteromonasand Marinomonas—Pseudoalteromonas atlanticaIAM12927^T, P. aurantiaNCIMB 2033^T, P. citreaATCC 29719^T, P. elyakoviiKMM 162^T, P. espejianaATCC 29659^T, P. piscicidaNCIMB 645^T, P. tetraodonisIAM 14160^T, Marinomonas communisATCC 27118^T, and M. vagaATCC 27119^T—showed that they contain glucose, galactose, galactosamine, glucosamine, fucose, rhamnose, mannose, heptose, 2-keto-3-deoxyoctonate (KDO), uronic acids, colitose (3,6-dideoxy-L-xylo-hexose), and 6-deoxy-L-talose. The carbohydrate composition of the antigenic polysaccharides (PSs) of P. elyakoviiKMM 162^Tand P. espejianaATCC 29659^Tdepended on the type and the concentration of carbohydrate substrates in the nutrient media. The molar proportion between rhamnose, glucose, and galactose (ca. 1 : 0.3 : 2) in the PS of P. elyakoviiKMM 162^Twas almost the same in the media lacking carbohydrates or containing glucose or galactose at a concentration of 1 g/l. At the same time, the molar proportion between fucose, glucose, galactose, galactosamine, and glucosamine (ca. 1 : 1 : 1 : 2 : 0.5) in the PS of P. espejianaATCC 29659^Tdepended on the presence and the concentration of carbohydrate substrates in the medium. A high concentration of glucose in the medium (30 g/l) brought about a rise in the content of glucose in PSs (9-fold for the PS of P. elyakoviiKMM 162^Tand 4.6-fold for the PS of P. espejianaATCC 29659^T) and led to a decrease in the content of other carbohydrates. The cultivation of these two strains at a lactose concentration of 30 g/l resulted in their PSs containing glucose and galactose in about equal proportions (ca. 1 : 1 in the case of P. espejianaATCC 29659^Tand ca. 2.1 : 1.7 in the case of P. elyakoviiKMM 162^T).     

The polysaccharide component of Pseudomonas pseudomallei slime

The polysaccharide component contained in the slime of the causative agent of melioidosis was obtained. This component was found to be a biopolymer, mainly of the carbohydrate nature, consisting of galactose, glucose, mannose, rhamnose and two unidentified carbohydrates. The slime polysaccharide component contained two thermostable and acid-resistant antigens. The action of alkali led to the loss of one of these antigens. Rabbit antisera to the preparations of the slime polysaccharide component with titers of 1:64 to 1:256, determined in the immunodiffusion test, were obtained. In the precipitation test the slime polysaccharide component reacted with antisera from sick experimental animals, thus confirming the suggestion of its secretion in the process of the development of melioidosis infection.     

Roles of RNA polymerase IV in gene silencing

Eukaryotes typically have three multi-subunit enzymes that decode the nuclear genome into RNA: DNA-dependent RNA polymerases I, II and III (Pol I, II and III). Remarkably, higher plants have five multi-subunit nuclear RNA polymerases: the ubiquitous Pol I, II and III, which are essential for viability; plus two non-essential polymerases, Pol IVa and Pol IVb, which specialize in small RNA-mediated gene silencing pathways. There are numerous examples of phenomena that require Pol IVa and/or Pol IVb, including RNA-directed DNA methylation of endogenous repetitive elements, silencing of transgenes, regulation of flowering-time genes, inducible regulation of adjacent gene pairs, and spreading of mobile silencing signals. Although biochemical details concerning Pol IV enzymatic activities are lacking, genetic evidence suggests several alternative models for how Pol IV might function.     

Chemical composition and serological analysis of the cell wall of Peptostreptococcus

Bahn, Arthur N. (Northwestern University, Chicago, Ill.), Patrick C. Y. Kung, and James A. Hayashi. Chemical composition and serological analysis of the cell wall of Peptostreptococcus. J. Bacteriol. 91:1672-1676. 1966.-Chemical and serological analyses were made of the cell wall of Peptostreptococcus to characterize taxonomically this genus of anaerobic streptococci. Cell wall hydrolysates of P. putridus strains 06 and 85, P. intermedius strains 11 and 87, and P. elsdenii strain B-159 were prepared, and the cell wall sugars were measured quantitatively by paper chromatography. Strain 85 contained only glucose, whereas strain 06 contained 93% glucose and 7% mannose. Strain 87 contained only rhamnose, and strain 11 contained approximately equal amounts of glucose and rhamnose. Strain B-159 differed from all the other strains in having a low (3.1%) content of total carbohydrate, consisting of rhamnose, galactose, and glucose. Quantitative amino acid analyses showed that the major amino compounds present in the cell wall were glutamic and aspartic acids, alanine, lysine, muramic acid, glucosamine, and galactosamine. Strains 06 and 85 possessed this complement of amino compounds, but strains 11 and 87 had relatively little aspartic acid. Strain B-159 was markedly different in having a high content of glycine and diaminopimelic acid, with only traces of lysine; it was the only strain in which teichoic acid was found. Serological analyses were made with the use of cell wall extracts as antigenic material and with homologous antisera, as well as streptococcal group antisera for groups A through S. The only strong agglutination was obtained between strain 87 antigen and group C antisera; weak agglutination was obtained with 87 against N, O, and K, and between strain 11 and groups E and F. All other antisera gave negative reactions. It is concluded that strain B-159 does not belong to the genus Peptostreptococcus, that strains 06 and 85 are members of P. putridus, and that strains 11 and 87 may be members of two different genera.     

Carbohydrate Composition of Variant-Specific Surface Antigen Glycoproteins from Trypanosoma brucei

SYNOPSIS. The carbohydrate of variant-specific surface antigen glycoproteins from bloodstream forms of 13 cloned variants of Trypanosoma brucei was analyzed by gas-liquid chromatography. The glycoproteins contained from 6 to 17% carbohydrate by weight, and all contained the same 4 sugars: mannose, galactose, glucose, and glucosamine (probably as N-acetyl-glucosamine). The glycoprotein from variant 048, strain 427 contained (±20%) 11 mannose, 4 galactose, 4 glucose, and 5 glucosamine residues/mole of glycoprotein (molecular weight 65,000). Glucose was an integral component of the glycoproteins, not dissociable by sodium dodecyl sulphate, 8 M urea, or 1 M acetic acid. Some of the glucose was dissociated by trichloroacetic acid. Most of the glycoproteins formed precipitin bands with concanavalin A in Ouchterlony double diffusion, but none formed such bands with wheat germ agglutinin or Ricinus communis lectin (molecular weight 120,000).     

Carbohydrate composition of variant-specific surface antigen glycoproteins from Trypanosoma brucei

The carbohydrate of variant-specific surface antigen glycoproteins from bloodstream forms of 13 cloned variants of Trypanosoma brucei was analyzed by gas-liquid chromatography. The glycoproteins contained from 6 to 17% carbohydrate by weight, and all contained the same 4 sugars: mannose, galactose, glucose, and glucosamine (probably as N-acetylglucosamine). The glycoprotein from variant 048, strain 427 contained (+20%) 11 mannose, 4 galactose, 4 glucose, and 5 glucosamine residues/mole of glycoprotein (molecular weight 65,000). Glucose was an intergral component of the glycoproteins, not dissociable by sodium dodecyl sulphate, 8 M urea, or 1 M acetic acid. Some of the glucose was dissociated by trichloroacetic acid. Most of the glycoproteins formed precipitin with concanavalin A in Ouchterlony double diffusion, but none formed such bands with wheat germ agglutinin or Ricinus communis lectin (molecular weight 120, 000).     

Studies on the structure of a phosphoglycoprotein from the parasitic protozoan Trypanosoma cruzi.

A glycoprotein (GP72) has been isolated from Trypanosoma cruzi and found to contain 41% protein, 49% carbohydrate and 10% phosphate. All phosphate was covalently attached to the carbohydrate which contained the following sugars: ribose, xylose, fucose, galactose, mannose, glucose and glucosamine. The carbohydrate side chains were linked to protein by fucose, xylose and N-acetylglucosamine; 50% of the total N-acetylglucosamine was involved in glycoprotein linkages. Two classes of carbohydrate side chains were detected. One class comprised 15% of the total carbohydrate and contained glucosamine, mannose and galactose; some of these chains were phosphorylated. The other class comprised 85% of the total carbohydrate and contained xylose, ribose, fucose, galactose, mannose, glucosamine and phosphate; these chains were antigenic and reacted with a monoclonal antibody with specificity for the whole glycoprotein.     

Capsular polysaccharides of nongroupable streptococci that cross-react with pneumococcal group 19

The cross-reactivity and chemical characterization of the nongroupable streptococcal and pneumococcal group 19 polysaccharides (PS) have been studied. Extensive cross-reactions were observed between capsular PSs of streptococcal strains 14636/74, 4907, 4731 and pneumococcal type 19F and 19A antisera. Streptococcal 14636/74 PS had an identical composition to that of pneumococcal 19F PS. Type 19F and 14636/74 PS were composed of equimolar amounts of rhamnose, glucose, N-acetyl mannosamine, and phosphorus. The capsular PS of strains 4731 and 4907 contained rhamnose, glucose, ribose, N-acetyl mannosamine, and N-acetyl glucosamine in different molar ratios. Extensive immunologic reactivity was observed between the 19F and 14636/74 PS, as determined by light scattering rate nephelometry, passive immune hemolysis, and precipitin reaction. There was an identity reaction by immunodiffusion between type 19F and 14636/74 PS when reacted with rabbit antiserum against either organism. Biochemical studies showed that strain 14636/74 was not a pneumococcus, because it was optochin resistant, was bile insoluble, did not possess the C-carbohydrate antigen common to all pneumococci, and produced neither pneumolysin nor IgA protease. Furthermore, it grew in comparatively simple media in contrast to the complex nutritional requirements of pneumococci. The 13C-NMR spectra of the 19F and 14636/74 PS were identical. These two capsular PS can, therefore, be considered identical.     

Characterization of lipopolysaccharides from Ralstonia solanacearum

Lipopolysaccharides (LPSs) from four strains of Ralstonia solanacearum belonging to biovar I (ICMP 6524, 8115, 5712, and 8169) were isolated and investigated. The structural components of the LPS molecule, such as lipid A, the core oligosaccharide, and O-specific polysaccharide (O-PS), were obtained after mild acid hydrolysis of the LPS preparations. In lipid A from all the LPS samples studied, 3-hydroxyhexadecanoic, 2-hydroxyhexadecanoic, tetradecanoic, and hexadecanoic fatty acids prevailed. The dominant monosaccharides of the core oligosaccharides of all of the strains studied were rhamnose, glucose, glucosamine, 2-keto-3-deoxyoctulosonic acid, and heptose. However, individual strains varied in the content of galactose, ribose, xylose, and arabinose. Three types of the O-PS structure were established, which differed in their configuration (alpha or beta), as well as in the type of the bond between glucosamine and rhamnose residues (1-->2 or 1-->3).     

Immunochemical studies on alpha-amylase. 3. Immunochemical relationships among amylases from various microorganisms

Sirishinha, Stitaya (University of Rochester School of Medicine and Dentistry, Rochester, N.Y.), and Peter Z. Allen. Immunochemical studies on alpha-amylase. III. Immunochemical relationships among amylases from various microorganisms. J. Bacteriol. 90:1120-1128. 1965.-Immunochemical relationships among amylases obtained from a selected group of microorganisms were examined, and a cross-reaction was detected between the alpha-amylases of Bacillus stearothermophilus and B. subtilis. Immunodiffusion and quantitative precipitin studies, as well as cross-neutralization tests, indicate that B. stearothermophilus alpha-amylase reacts with a portion of antibody present in antisera to crystalline B. subtilis alpha-amylase. Amylases from these two species thus have some aspects of structure in common. Limited data obtained by immunodiffusion suggest that groupings which confer cross-reactivity to the B. stearothermophilus enzyme are lost after exposure to mercaptoethanol in the presence of ethylenediamine-tetraacetate, followed by treatment with iodoacetamide. With the antisera employed and within the concentration range examined, no immunochemical cross-reaction was observed among amylases from Aspergillus oryzae, B. subtilis, B. polymyxa, B. macerans, Pseudomonas saccharophila, and Euglena sanguinis. Immunoelectrophoresis of partially purified B. stearothermophilus alpha-amylase by use of antiserum to the crude enzyme, together with localization of amylase activity in immunoelectrophoretic plates, suggests that B. stearothermophilus alpha-amylase is antigenic in the rabbit.     

Serological Relationships of Type I Antigens of Group B Streptococci

Some of the complex antigenic relationships of type I group B streptococci from various clinical sources were defined by means of immunodiffusion, absorption, and precipitin tests. Three predominant types are described: Ia, Ib, and Ii. Methods for preparing antisera for differentiating type I strains are presented.     

Anti-glycosyl antibodies: Preparation and characterization of rabbit anti-galactose and anti-lactose antibodies

Anti-galactose and anti-lactose antibodies have been isolated from the antisera of rabbits immunized with non-viable cells of $\text{Streptococcus}$$\text{faecalis}$, strain N containing an antigenic diheteroglycan of glucose and galactose in the cell wall. The anti-galactose antibodies are specific for the galactosyl moiety while the anti-lactose antibodies are specific for the lactosyl moiety of the diheteroglycan. Hapten inhibitions with galactose and lactose, the sedimentation constant, the immunoglobulin type, the carbohydrate content, the electrophoretic mobility and the amino acid composition have been determined for the two new types of anti-glycosyl antibodies.     

Immunological cross-reactivity between chicken slow skeletal and ventricular muscle myosin

Antibodies elicited in rabbits against chicken slow skeletal anterior latissimus dorsi and ventricular myosin were analyzed by double immunodiffusion for their ability to react with homologous and heterologous antigen at different stages of immunization (1--12 months). Each anti myosin antiserum formed a single, strong precipitin line with its immunogen after short time of immunization. This reaction was specific for myosin heavy chains as determined by GEDELISA (gel electrophoresis derived enzyme lined immunosorbent assay) test. In rabbits injected with ventricular myosin after long time of immunization a second, fainter precipitin line has generally been observed. The antigenic determinants responsible for this precipitin line have been localized on the light myosin subunits. By comparing the two types of anti myosin antisera with heterologous antigen we have obtained evidence for partial immunological cross-reactivity between slow skeletal and ventricular muscle myosins. In particular, all anti ventricular myosin antisera displayed a marked immunological reactivity with anterior latissimus dorsi myosin whereas most of anti anterior latissimus dorsi myosin antisera showed absence of reciprocity. By means of immunofluorescence and immunoabsorption techniques both common and unique slow skeletal and ventricular antigenic determinants have been demonstrated.     

Ampicillin-resistant mutants of Escherichia coli K-12 with lipopolysaccharide alterations affecting mating ability and susceptibility to sex-specific bacteriophages

Mutations from moderate (class I) to high (class III) ampicillin resistance in a male and a female strain of Escherichia coli K-12 have been found to be accompanied by surface alterations, first demonstrated as hindrance in the formation of mating pairs. These changes have now been studied with the ribonucleic acid phage MS2, and especially with the "female-specific" phage phiW. Several class III mutations in male and female strains were found to make the cells susceptible to phage phiW and to reduce their abilities to form mating pairs. Spontaneous phage phiW-resistant mutants isolated from class III strains were found also to have acquired changes in ampicillin resistance and ability to form mating pairs. One mutant had reverted to parental class I type in all three properties. Lipopolysaccharides (LPS) prepared from phiW-sensitive class III strains inactivated the phage in vitro, whereas LPS from phage-resistant strains had no effect. Carbohydrate analyses of LPS preparations showed that two class III mutants, compared to their parental strains, had lost significant parts of the rhamnose, galactose, and glucose from the LPS. One of the phage phiW-resistant mutants showed a partial restoration of its carbohydrate composition. Other phiW-resistant mutants showed, instead, further losses of carbohydrates in their LPS. It is suggested that genes exist which simultaneously mediate a female-specific mating site, ampicillin resistance, and the receptors for phage phiW.     

Multiple polysaccharide antigens of group B streptococcus, type Ia: emphasis on a sialic acid type-specific polysaccharide.

Group B streptococci, type Ia (strain 090/14/4), were subjected to sequential extraction procedures and the various extracts were fractionated by a combination of DEAE-cellulose chromatography and gel-filtration. Three major antigens were isolated: the conventional group-specific and type-specific carbohydrates and an acidic polysaccharide consisting of galactose, glucose, glucosamine, and sialic acid. Immunochemical data suggest that the acidic antigen, with the exception of the immunodominant sialic acid, is structurally similar to the conventional pH 2.0 extracted type-specific antigen. Removal of the terminal sialic acid residues from the acidic antigen by mild acid hydrolysis resulted in a residual polymer which was chemically and immunologically analogous to the type-specific carbohydrate. Precipitin analysis focused attention to the cross-reactivity between thia acidic antigen and anti-types Ib, Ic, and III sera.     

Cell wall composition of chlorococcal algae

The cell walls of representatives of the genera Chlorella, Monoraphidium, Ankistrodesmus and Scenedesmus contained 24–74% neutral sugars, 1–24% uronic acids, 2–16% protein and 0–15% glucosamine. Two types of cell walls could be discerned containing as main sugars either rhamnose and galactose or mannose and glucose with a lack of galactose.