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Biomedical subjects

J Adler

Publications and source records attributed to J Adler.

At least 289 records · Page 16Linked to original sources
PubMed1971

Purification of intact flagella from Escherichia coli and Bacillus subtilis.

A procedure is described for the purification of bacterial flagella in the form of a filament-hook-basal body complex (intact flagella) free from detectable cell wall, membrane, or cytoplasmic material. Spheroplasts produced with lysozyme and ethylenediaminetetraacetic acid were lysed with Triton X-100, and the flagella were purified by (NH(4))(2)SO(4) precipitation, differential centrifugation, and CsCl gradient centrifugation. As much as 40% of the flagella were recovered, and they contained about one basal body per 4 to 6 mum of flagella. The same procedure developed for Escherichia coli was also successful for purifying intact flagella from Bacillus subtilis.

Bacillus subtilis↗
PubMed1971

Fine structure and isolation of the hook-basal body complex of flagella from Escherichia coli and Bacillus subtilis.

The hook-basal body complex comprising the basal end of purified intact flagella from Escherichia coli and Bacillus subtilis was studied in detail with an electron microscope. The E. coli hook can be described as having five or six concentric helical coils. The basal body from E. coli is 27 nm in length and consists of four rings, 22.5 nm in diameter, arranged in two pairs and mounted on a rod. The top pair of rings is connected near their periphery, resembling a closed cylinder. In B. subtilis the basal body looks like that from E. coli, except that the top pair of rings is missing. Hook-basal body complexes from both organisms could be isolated by dissociating the filaments with either urea or acid. Based on our results, two types of basal body structures are proposed, as exemplified by E. coli and B. subtilis, which directly reflect the structure of the gram-negative and gram-positive cell envelopes.

Acetates↗
PubMed1971

Attachment of flagellar basal bodies to the cell envelope: specific attachment to the outer, lipopolysaccharide membrane and the cyoplasmic membrane.

A procedure is described for the purification of the Escherichia coli outer membrane (lipopolysaccharide or L membrane) with flagella still attached. The resulting lipopolysaccharide membrane was in the form of vesicles that had a trilaminar structure in thin section and contained about 55% lipopolysaccharide and 45% protein. T2 or T4 phage preadsorbed to E. coli were found attached to the purified lipopolysaccharide membrane. Flagella were bound to the purified lipopolysaccharide membrane specifically at the basal body ring closest to the hook (the L ring). The cytoplasmic membrane in preparations from osmotically lysed E. coli spheroplasts or Bacillus subtilis protoplasts was specifically attached to flagella at the basal body ring farthest from the hook (the M ring). In the E. coli preparation, lipopolysaccharide membrane was also present and was attached to the L ring. From these data and a knowledge of the structure and dimensions of the E. coli flagellar basal body and cell envelope, a model for flagellar attachment is deduced.

Adenosine Triphosphatases↗
PubMed1969

Escherichia coli mutants defective in chemotaxis toward specific chemicals.

Mutants of Escherichia coli K12 have been found which fail to carry out chemotaxis toward certain chemicals only. One mutant exhibits greatly reduced chemotaxis toward L-serine but has no detectable defect either in uptake or in oxidative metabolism of that compound. Another mutant is not attracted to D-galactose and certain related sugars. There is a correlation between the galactose chemotaxis defect and a defect in galactose uptake, perhaps indicating a common component for chemotaxis and uptake systems. The results are discussed in terms of a model for chemotaxis in which attractants are detected by specific "chemoreceptors."

Chemotaxis↗
PubMed1969

Location of genes for motility and chemotaxis on the Escherichia coli genetic map.

In Escherichia coli, the following genes are involved in motility and chemotaxis. The H gene is the structural gene for flagellin. Mutation in the mot gene results in paralysis of the flagella, and mutation in the fla genes leads to an absence of flagella. The cheA, cheB, and cheC genes are required for chemotaxis. The chromosomal location of these genes has now been determined. The majority are clustered in a small region around uvrC, between his and aroD, in the order his-cheC-H-uvrC-mot-cheA-cheB-aroD. The fla genes are located in the same region, and also between trp and gal. The results indicate that many of the genes are homologous to those which have been studied in Salmonella typhimurium.

Cell Biology↗
PubMed1967

Nonchemotactic mutants of Escherichia coli.

We have isolated 40 mutants of Escherichia coli which are nonchemotactic as judged by their failure to swarm on semisolid tryptone plates or to make bands in capillary tubes containing tryptone broth. All the mutants have normal flagella, a fact shown by their shape and reaction with antiflagella serum. All are fully motile under the microscope and all are sensitive to the phage chi. Unlike its parent, one of the mutants, studied in greater detail, failed to show chemotaxis toward oxygen, glucose, serine, threonine, or aspartic acid. The failure to exhibit chemotaxis does not result from a failure to use the chemicals. The swimming of this mutant was shown to be random. The growth rate was normal under several conditions, and the growth requirements were unchanged.

Aspartic Acid↗
PubMed1967

Purification and chemistry of bacteriophage chi.

From a stock of varkappa phage grown on Salmonella, a host-range mutant which attacks Escherichia coli was isolated. As in the case of Salmonella, only motile strains of E. coli are sensitive to varkappa. The phage shows an eclipse period of 35 min and a minimal latent period of 52 min. The adsorption rate constant is 3 x 10(-9) ml/min. Adsorption shows a marked dependence on temperature. Bacteriophage varkappa was purified by differential centrifugation and CsCl density gradient centrifugation. It contains deoxyribonucleic acid (DNA) which is double-stranded. The DNA has a molecular weight of 42 million and a guanine plus cytosine content of 57%. Of 68 molecules of DNA inspected, 7 were circular. The phage particle weight is about 90 million.

Adsorption↗
PubMed1967

How bacteriophage chi attacks motile bacteria.

Bacteriophage chi attaches to the filament of a bacterial flagellum by means of a tail fiber, but the ultimate receptor site for the phage is located at the base of the bacterial flagellum. Here, the phage injects its deoxyribonucleic acid into the bacterium, leaving the empty phage attached at the base. It is suggested that chi slides along the filament of the flagellum to the base, owing to the movement of the flagellum. The role of motility would thus be to provide for rapid adsorption of the phage by guiding the phage to the adsorption sites at the bases of the flagella. Bacteria whose motility has been strongly inhibited by cold or anaerobic conditions still adsorb chi at the filaments and bases of flagella if a high multiplicity is used. This indicates that direct collisions with the bases may also be possible. Bacteria must be flagellated in order for chi to attach, but only a short flagellum, perhaps only the flagellar base, is necessary.

Adsorption↗
PubMed1966

Chemotaxis in bacteria.

Motile Escherichia coli placed at one end of a capillary tube containing an energy source and oxygen migrate out into the tube in one or two bands, which are clearly visible to the naked eye and can also be demonstrated by photography, microscopy, and densitometry and by assaying for bacteria throughout the tube. The formation of two bands is not due to heterogeneity among the bacteria, since the bacteria in each band, when reused, will form two more bands. If an anaerobically utilizable energy source such as galactose is present in excess over the oxygen, the first band consumes all the oxygen and a part of the sugar and the second band uses the residual sugar anaerobically. On the other hand, if oxygen is present in excess over the sugar, the first band oxidizes all the sugar and leaves behind unused oxygen, and the second band uses up the residual oxygen to oxidize an endogenous energy source. The essence of the matter is that the bacteria create a gradient of oxygen or of an energy source, and then they move preferentially in the direction of the higher concentration of the chemical. As a consequence, bands of bacteria (or rings of bacteria in the case of agar plates) form and move out. These results show that E. coli is chemotactic toward oxygen and energy sources such as galactose, glucose, aspartic acid, threonine, or serine. The full repertoire of chemotactic responses by E. coli is no doubt greater than this, and a more complete list remains to be compiled. The studies reported here demonstrate that chemotaxis allows bacteria to find that environment which provides them with the greatest supply of energy. It is clearly an advantage for bacteria to be able to carry out chemotaxis, since by this means they can avoid unfavorable conditions and seek optimum surroundings. Finally, it is necessary to acknowledge the pioneering work of Englemann, Pfeffer, and the other late-19thcentury biologists who discovered chemotaxis in bacteria, and to point out that the studies reported here fully confirm the earlier reports of Beijerinck (4) and Sherris and his collaborators (5,6) on a band of bacteria chemotactic toward oxygen. By using a chemically defined medium instead of a complex broth, it has been possible to study this band more closely and to demonstrate in addition the occurrence of a second band of bacteria chemotactic toward an energy source. Beijerinck (4) did, in fact, sometimes observe a second band, but he did not offer an explanation for it.

Amino Acids↗