SEARCH · PubMed Health

Results for “FLAGELLA”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 307 records · Page 17Linked to original sources

PubMed1987

Localization and stoichiometry of hook-associated proteins within Salmonella typhimurium flagella.

The localization of hook-associated proteins (HAP1, HAP2, and HAP3) in Salmonella typhimurium flagella was studied by using specific antibodies together with a second antibody conjugated with colloidal gold. HAP1 and HAP3 were localized at the hook-filament junction, as has been suggested previously. HAP2, however, was localized at the filament tip. This finding supports the idea that HAP2 acts to induce polymerization of endogenous flagellin at the filament tip, and HAP1 and HAP3 are junction proteins to connect hook with filament. Analysis of the protein composition of short flagella from a mutant indicated that a single flagellum contains about 10 to 20 HAP1, 10 to 20 HAP2, and 10 to 40 HAP3 molecules.

Antibodies↗

PubMed1987

Formation of flagella lacking outer rings by flaM, flaU, and flaY mutants of Escherichia coli.

Among flagellar mutants of Escherichia coli, flaM or flaU mutants form basal bodies lacking the outer P and L rings, whereas flaY mutants predominantly form basal bodies lacking the L ring. In these mutants, hooks and filaments are occasionally assembled onto these incomplete basal bodies. When the hook protein gene, flaFV, of Salmonella typhimurium was cloned on the multicopy plasmid pBR322 and introduced into these mutants, the efficiency with which cells assembled hooks and filaments onto the incomplete basal bodies increased significantly. Such cells formed characteristic dotted swarms on semisolid plates, indicating that cells carrying flagella without the outer rings are weakly motile because of poor function of their flagella, a low flagellar number per cell, or both of these defects. FlaV mutants also produced incomplete basal bodies lacking the outer rings, but assembly of hooks and filaments did not occur in these mutants even after introduction of the plasmid carrying flaFV of S. typhimurium. The failure in the case of flaV mutants was attributed to their inability to modify the rod tip to the structure competent for assembly of hook protein.

Bacterial Proteins↗

PubMed1988

Biochemical and cytological analysis of the complex periplasmic flagella from Spirochaeta aurantia.

The periplasmic flagella of Spirochaeta aurantia were isolated and were found to be ultrastructurally and biochemically complex. Generally, flagellar filaments were 18 to 20 nm in diameter and appeared to consist of an 11 to 13-nm-wide inner region and an outer layer. The hook-basal body region consisted of two closely apposed disks connected to a hook by a rod. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified flagella together with a Western blot analysis of a motility mutant that produces hooks and basal bodies but not flagellar filaments revealed that the filaments were composed of three major polypeptides of 37,500, 34,000, and 31,500 apparent molecular weight (37.5K, 34K, and 31.5K polypeptides) and three minor polypeptides of 36,000, 33,000, and 32,000 apparent molecular weight (36K, 33K, and 32K polypeptides). Purified hook-basal body preparations were greatly enriched in three polypeptides in the range of 62,000 to 66,000 apparent molecular weight. Immunogold labeling experiments with a monoclonal antibody specific for the 37.5K flagellin and one that reacts with an epitope common to the 36K, 34K, 33K, 32K, and 31.5K flagellins revealed that the 37.5K major polypeptide was a component of the outer layer, whereas one or more of the other polypeptides constituted the core.

Antigens, Bacterial↗

PubMed1993

Mechanism of adverse conditions causing lack of flagella in Escherichia coli.

Escherichia coli lacks flagella when grown in tryptone broth in the presence of various adverse conditions (C. Li, C. J. Louise, W. Shi, and J. Adler, J. Bacteriol. 175:2229-2235, 1993). Now, the synthesis, rather than the degradation, of flagellin was shown to be inhibited. Studies of transcriptional fusions of flagellar operons to the lacZ gene revealed that transcription of the flagellar genes was reduced in cells grown under these adverse conditions. Increasing gene dosage of the flhD operon by a plasmid partially suppressed the nonflagellation caused by some adverse conditions. The signal which shuts off the synthesis of flagella under adverse conditions remains to be discovered. This shutting-off process does not result from catabolite repression or from signals from the chemotaxis system.

Chemotaxis↗

PubMed1996

Effect of viscosity on swimming by the lateral and polar flagella of Vibrio alginolyticus.

By using mutants of Vibrio alginolyticus with only a polar flagellum (Pof+ Laf-) or only lateral flagella (Pof- Laf+), we examined the relationship between swimming speed and the viscosity of the medium for each flagellar system. Pof+ Laf- cells could not swim in the high-viscosity environment (ca. 200 cP) in which Pof- Laf+ cells swam at 20 microns/s. The Pof- Laf+ cells swam at about 20 microns/s at normal viscosity (1 cP) without the viscous agent, and the speed increased to 40 microns/s at about 5 cP and then decreased gradually as the viscosity was increased further. These results show the functional difference between polar and lateral flagella in viscous environments.

Cell Movement↗

PubMed2003

Flagella and Motility in Actinobacillus pleuropneumoniae.

Actinobacillus pleuropneumoniae has been considered nonmotile and nonflagellate. In this work, it is demonstrated that A. pleuropneumoniae produces flagella composed of a 65-kDa protein with an N-terminal amino acid sequence that shows 100% identity with those of Escherichia coli, Salmonella, and Shigella flagellins. The DNA sequence obtained through PCR of the fliC gene in A. pleuropneumoniae showed considerable identity (93%) in its 5' and 3' ends with the DNA sequences of corresponding genes in E. coli, Salmonella enterica, and Shigella spp. The motility of A. pleuropneumoniae was observed in tryptic soy or brain heart infusion soft agar media, and it is influenced by temperature. Flagella and motility may be involved in the survival and pathogenesis of A. pleuropneumoniae in pigs.

Actinobacillus Infections↗

PubMed1967

Basal organelles of bacterial flagella.

Liberated by enzymatic lysis of the cells, the flagella of Rhodospirillum rubrum, R. molischianum, and R. fulvum all have a similar structure. The hook at the base of the flagellum is connected by a short, narrow collar to a paired disc in the basal organelle. This paired disc is in turn connected to a second paired disc. The disposition of flagella to which fragments of the cell membrane still adhere suggests that the narrow collar at the base of the hook traverses both the wall and the membrane, and that the upper pair of discs in the basal organelle lies just beneath the surface of the membrane.

Bacteriolysis↗

PubMed1968

Use of radioactive antibodies for characterizing antigens and application to the study of flagella synthesis.

A simple rapid immunochemical procedure has been developed which provides information about the qualitative and quantitative nature of antigens. It involves the use of purified radioactive ((125)I-labeled) antibodies. The amount of antibody bound to the antigen is determined by filtering the mixture through diethylaminoethyl (DEAE)-cellulose paper. All of the antigen, as well as the antibody complexed with it, is trapped on the paper, whereas free antibody is removed by repeated washing. This technique has been applied to the study of three immune systems, bovine serum albumin, Escherichia coli tryptophan synthetase B protein, and Bacillus subtilis flagella. The results obtained by the DEAE-antibody binding technique were comparable, in terms of sensitivity, specificity, and accuracy, to data obtained by microcomplement fixation and precipitin methods. The assay was used to measure the kinetics of flagella regeneration in B. subtilis.

Antibodies↗

PubMed1969

Antigenic heterology between flagellin and flagella of Bacillus subtilis.

The antigenic properties of Bacillus subtilis flagella and periodate-treated flagellin were compared by using immobilization-inhibition and diethylaminoethyl cellulose binding assay. The results suggest that there are both unique and cross-reactive antibodies elicited by B. subtilis flagella and flagellin.

Antigens↗

PubMed1969

Mechanical removal of F pili, type I pili, and flagella from Hfr and RTF donor cells and the kinetics of their reappearance.

The effect of mechanical agitation (blending) on the removal of F pili, type I pili, and flagella from Hfr (high-frequency recombinant) and resistance transfer factor (RTF) fi(+)Escherichia coli cells was studied by electron microscopy. The reduction in number and length of appendages was measured as a function of blendor speed under standard conditions of temperature, medium, cell density, and blendor configuration. F pili and flagella were removed within the same narrow range of blendor speeds. Type I pili were removed within a higher and broader range of speeds. The speed which reduced the average length of type I pili to 50% was 3.5 times the speed which reduced the average length of F pili to 50%. None of the speeds employed inhibited cell growth, viability, or the ability to produce cell appendages. The kinetics of reappearance of F pili and type I pili after removal by blending were also different. F pili grew out very rapidly, reaching 50% of their full length in 30 sec and their full length in 4 to 5 min. The number of attached F pili per cell also increased rapidly, reaching a constant value in 4 to 5 min. After 5 min, F pilus lengths were distributed around a modal value of about 1.2 mum, and the numbers of F pili per cell were distributed according to a Poisson distribution, with an average of 1.0 per cell. These reappearance kinetics, length distributions, and number distributions are consistent with a model of F-pilus outgrowth in which new F pili appear at random locations on the cell surface at an average rate of about once every 4 min, grow to their characteristic length in about 4 min, and then separate from the cell. F pili which had separated could absorb to the cells, leading to the presence of two classes of F pili on cells: those in the process of natural out-growth and those attached by absorption. Type I pili increased in length much more slowly than did F pili, although the fraction of cells having visible type I pili increased very rapidly after blending because of the large number of type I pili per cell. The fraction of flagellated cells increased even more slowly, reaching only 30% of the unblended fraction in 30 min. The application of blending spectra and reappearance kinetics to the identification of cell functions with surface structures is discussed.

Coliphages↗

PubMed1988

Characterization of flagella purified from enterohemorrhagic, vero-cytotoxin-producing Escherichia coli serotype O157:H7.

Escherichia coli of the serotype O157:H7 has recently been isolated in human fecal specimens in association with sporadic cases and outbreaks of hemorrhagic colitis and with the hemolytic uremic syndrome. The aim of this study was to characterize the flagellin protein subunit constituents of flagellar filaments from E. coli O157:H7 strain CL-56. Flagellin isolated from a reference Salmonella enteritidis strain was used for comparison. Flagella were dissociated by incubation of bacteria under acidic conditions, centrifugation, and differential ammonium sulfate precipitation. Reconstituted flagellar filaments were demonstrated by three complementary methods: transmission electron microscopy, antigenic reactivity with H7 antiserum by a dot blot immunoassay, and immunogold localization of antiserum raised to the purified antigen to intact flagella on whole E. coli O157:H7. On sodium dodecyl sulfate-polyacrylamide gels flagellin proteins from E. coli O157:H7 demonstrated an apparent Mr of 66,000. The isoelectric point of E. coli O157:H7 flagellin was 5.42. By immunoblotting, H7 flagellin proteins were shown to be immunogenic. They induced a systemic immune response both in rabbits challenged with whole bacteria and in a human previously infected with E. coli O157:H7.

Actin Cytoskeleton↗

PubMed1989

Staining bacterial flagella easily.

A wet-mount technique for staining bacterial flagella is highly successful when a stable stain and regular slides and cover slips are used. Although not producing a permanent mount, the technique is simple for routine use when the number and arrangement of flagella are critical in identifying species of motile bacteria.

Alcaligenes↗

PubMed1990

Characterization of flagella of Clostridium difficile and their role in serogrouping reactions.

Slide agglutination with rabbit antisera allows the differentiation of 10 serogroups of Clostridium difficile, namely, A, B, C, D, F, G, H, I, K, and X. Each serogroup displays a specific protein profile in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, except for A, which displays 12 different protein profiles (A1 to A12). In the present work, electron microscopy revealed the presence of uniformly distributed flagella in the reference strains of serogroups G and K and in all strains representative of the 12 subgroups within serogroup purified by differential centrifugation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of these preparations revealed one distinct band with an apparent molecular mass of approximately 39 kilodaltons. Antiserum was prepared by immunizing a rabbit with the serogroup A flagellin, which had been eluted from the gel. In immunoblotting, this antiserum cross-reacted with the flagellin of the other strains. When the cells were deflagellated by a short sonication, the cross-reactions observed by slide agglutination with A, G, and K antisera were suppressed. Similarly, shearing of flagella allowed specific slide agglutination of the 12 subgroups of serogroup A.

Agglutination Tests↗

PubMed1997

DNA elements regulating alpha1-tubulin gene induction during regeneration of eukaryotic flagella.

Eukaryotic flagella are complex organelles composed of more than 200 polypeptides. Little is known about the regulatory mechanisms governing synthesis of the flagellar protein subunits and their assembly into this complex organelle. The unicellular green alga Chlamydomonas reinhardtii is the premier experimental model system for studying such cellular processes. When acid shocked, C. reinhardtii excises its flagella, rapidly and coordinately activates transcription of a set of flagellar genes, and ultimately regenerates a new flagellar pair. To define functionally the regulatory sequences that govern induction of the set of genes after acid shock, we analyzed the alpha1-tubulin gene promoter. To simplify transcriptional analysis in vivo, we inserted the selectable marker gene ARG7 on the same plasmid with a tagged alpha1-tubulin gene and stably introduced it into C. reinhardtii cells. By deletion of various sequences, two promoter regions (-176 to -122 and -85 to -16) were identified as important for induction of the tagged alpha1-tubulin gene. Deleting the region between -176 and -122 from the transcription start site resulted in an induction level which was only 45 to 70% of that of the resident gene. Deleting the region upstream of -56 resulted in a complete loss of inducibility without affecting basal expression. The alpha1-tubulin promoter region from -85 to -16 conferred partial acid shock inducibility to an arylsulfatase (ARS) reporter gene. These results show that induction of the alpha1-tubulin gene after acid shock is a complex response that requires diverse sequence elements.

Animals↗

PubMed1980

Serological relationships among flagella of Erwinia carotovor var. atroseptica and some E. carotovora var. carotovora serogorups.

The 2 Erwinia carotovora var. atroseptica serogroups and 2 out of 16 E. carotovora var. carotovora serogroups previously established on the basis of diffusible somatic antigens were shown to be serologically related by agglutination procedures using whole cells. The common agglutinating antigen in serogroups I, III, V, and XVIII was heat labile and identified as the bacterial flagella by the fluorescent antibody staining procedure. A few strains in serogorups I and III apparently lacked flagella altogether. Fluorescent antibody staining of whole cells also confirmed that the cell wall antigens of serogroups I, II, and XVIII were related and that the cell wall antigens of serogroups III and V were not related to each other or to the other serogroups.

Agglutination Tests↗

PubMed1991

Antigenic and immunogenic activity of flagella and fimbriae preparations from uropathogenic Proteus mirabilis.

The antigenic and immunogenic activities of fimbriae and flagella from three uropathogenic strains of Proteus mirabilis were compared. Flagella were obtained by mechanical treatment and fimbriae were isolated from cells by heat shock, ammonium sulfate precipitation, sodium deoxycholate and urea treatment, and gel filtration. Both preparations inoculated to mice demonstrated high antigenicity. Titers up to 1:80,000 were determined by an enzyme-linked immunosorbent assay either against the homologous or heterologous strains. When immunized mice were challenged with homologous or heterologous hematogenous infecting doses, a good cross protection was achieved only when fimbriae were used as antigens. Cross-reactivity found between the three fimbriae antisera, and the presence of common proteins in sodium dodecyl sulfate polyacrylamide gel electrophoresis patterns of fimbriae, should validate the study of these proteins to determine the existence of a shared adhesin.

Animals↗

PubMed2004

Association of sperm protein 17 with A-kinase anchoring protein 3 in flagella.

BACKGROUND: Sperm protein 17 (Sp17) is a three-domain protein that contains: 1) a highly conserved N-terminal domain that is 45% identical to the human type II alpha regulatory subunit (RII alpha) of protein kinase A (PKA); 2) a central sulphated carbohydrate-binding domain; and 3) a C-terminal Ca++/calmodulin (CaM) binding domain. Although Sp17 was originally discovered and characterized in spermatozoa, its mRNA has now been found in a variety of normal mouse and human tissues. However, Sp17 protein is found predominantly in spermatozoa, cilia and human neoplastic cell lines. This study demonstrates that Sp17 from spermatozoa binds A-kinase anchoring protein 3 (AKAP3), confirming the functionality of the N-terminal domain. METHODS: In this study in vitro precipitation and immunolocalization demonstrate that Sp17 binds to AKAP3 (AKAP110) in spermatozoa. RESULTS: Sp17 is present in the head and tail of spermatozoa, in the tail it is in the fibrous sheath, which contains AKAP3 and AKAP4. Recombinant AKAP3 and AKAP4 RII binding domains were synthesized as glutathione S-transferase (GST) fusion proteins immobilized on glutathione-agarose resin and added to CHAPS extracts of human spermatozoa. Western blots of bound and eluted proteins probed with anti-Sp17 revealed that AKAP3 bound and precipitated a significant level of Sp17 while AKAP4 did not. AKAP4 binds AKAP3 and expression of AKAP3 is reduced in AKAP4 knockout sperm, therefore we tested AKAP4 knockout spermatozoa for Sp17 and found that there was a reduction in the amount of Sp17 expressed when compared to wild type spermatozoa. Co-localization of AKAP3 and Sp17 by immunofluorescence was demonstrated along the length of the principal piece of the flagella. CONCLUSIONS: As predicted by its N-terminal domain that is 45% identical to the human RIIalpha of PKA, Sp17 from spermatozoa binds the RII binding domain of AKAP3 along the length of the flagella.

A Kinase Anchor Proteins↗

PubMed1994

Kinesin-related proteins in eukaryotic flagella.

To identify kinesin-related proteins that are important for ciliary and eukaryotic flagellar functions, we used affinity-purified, polyclonal antibodies to synthetic peptides corresponding to conserved sequences in the motor domain of kinesin (Sawin et al. (1992) J. Cell Sci. 101, 303-313). Using immunoblot analysis, two antibodies to distinct sequences (LNLVDLAGSE, 'LAGSE' and, HIPYRESKLT, 'HIPYR') reveal a family of proteins in flagella and axonemes isolated from Chlamydomonas. Similar analysis of axonemes from mutant Chlamydomonas strains or fractionated axonemes indicates that none of the immunoreactive proteins are associated with dynein arm or spoke structures. In contrast, one protein, approximately 110 kDa, is reduced in axonemes from mutant strains defective in the central pair apparatus. Immunoreactive proteins with masses of 96 and 97 kDa (the '97 kDa' proteins) are selectively solubilized from isolated axonemes in 10 mM ATP. The 97 kDa proteins co-sediment in sucrose gradients at about 9 S and bind to axonemes or purified microtubules in a nucleotide-dependent fashion characteristic of kinesin. These results reveal that flagella contain kinesin-related proteins, which may be involved in axonemal central pair function and flagellar motility, or directed transport involved in morphogenesis or mating responses in Chlamydomonas.

Adenosine Triphosphate↗