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At least 19 recordsLinked to original sources
PubMed1976

Amino acid sequence of flagellin of Bacillus subtilis 168. II. Tryptic peptides from maleylated flagellin.

Fifteen pure peptides, ranging in length from 2 to 141 residues, were isolated from the tryptic digest of maleylated flagellin. Studies of these peptides in conjunction with the data on the eight cyanogen bromide peptides described earlier (Chang, J. Y., DeLange, R. J., Shaper, J. H., and Glazer, A. N. (1976) J. Biol. Chem. 251, 695-700) established the order of three of the CNBr peptides at the NH2 terminus and of three at the COOH terminus of flagellin.

Amino Acid Sequence↗
PubMed1977

Regulation of flagellin synthesis in the cell cycle of caulobacter: dependence on DNA replication.

Synthesis of the two filament proteins (flagellin A and flagellin B) of the Caulobacter creascentus flagellum was measured during the cell cycle. Synchronous cells were pulse-labeled with 36S-methionine, and flagellin proteins were isolated from crude extracts by radioimmune precipitation. The results showed that both proteins are maximally induced during the G2 period and that their induction requires de novo transcription. Flagellin A, however, continues to be made in the progeny swarmer cells after flagellin B synthesis has stopped. This discoordination in flagellin A and B synthesis and the relative abundance of the two proteins may result in part from the longer functional half-life of the flagellin A messenger RNA. Analysis of temperature-sensitive DNA chain elongation mutants suggests that the periodicity of flagellin A and B synthesis in the cell cycle is controlled by a late cell cycle event, presumably the completion of chromosome replication.

Bacteria↗
PubMed1977

Comparison of the flagellins from different flagellar morphotypes of Escherichia coli.

The molecular weights of the flagellins of 13 strains of Escherichia coli, each with a different H antigen, were estimated using polyacrylamide gel electrophoresis. In each case only one major polypeptide was demonstrated, although some strains possessed apparently sheathed flagella. Considerable differences in the molecular weight of flagellin accompanied the previously described structural differences between flagella from strains with different H antigens. The relationship between flagellar diameter and the molecular weight of the corresponding flagellins was similar for both unsheathed and apparently sheathed flagella. Crosss-polymerization occurred between seed consisting of fragment of unsheathed flagella and flagellin solution from apparently sheathed flagella and vice versa. Co-polymerization of flagellin from unsheathed flagella and flagellin from apparently sheathed flagella was also demonstrated. These polymerization experiments indicate that the assembly pattern of flagellin molecules is probably the same in all E. coli flagella. The above and other evidence suggests that there is no true sheath, but that the differences in flagellar surface structure between different E. coli flagella are the result of differences in the superficial parts of the flagellin molecules.

Antigens, Bacterial↗
PubMed1978

Immunogenic capacity of ribosomes of Salmonella typhi interfered with a flagellin-like material contaminant.

The double-immunodiffusion technique and sodium dodecyl sulfate-polyacrylamide electrophoresis were used to demonstrate the presence of flagellin-like material strongly attached to ribosomes of Salmonella typhi Ty 2. This flagellin-like material contaminating the ribosome preparation interferes with the induction of antiribosome serum promoting the formation of antisera reacting either only with flagellin or in some cases with flagellin and ribosomes, but giving a very weak reaction with the latter. The interference is also observed when purified ribosomes from a nonflagellated mutant of S. typhi (S. typhi O-901) mixed with purified S. typhi Ty 2 flagellin are utilized as antigens. The antiribosome sera obtained with ribosomes from S. typhi O-901 have a considerably higher titer than those that are interfered with. These sera were able to react with ribosomes obtained from several related species and did not react with flagella-derived flagellin of S. typhi Ty 2.

Antigens, Bacterial↗
PubMed1979

M467: a murine IgA myeloma protein that binds a bacterial protein. I. Recognition of common antigenic determinants on Salmonella flagellins.

We have studied the binding of M467, an IgA murine myeloma protein, to flagellin from seven species of Salmonella. It was found that M467 was reacting with antigenic determinants that were common to all the flagellins studied. These determinants were not related to serotypic antigens. Electronmicrographs of unreduced M467 showed a variety of polymeric species bound to flagella in a manner that could produce immobilization as well as agglutination and precipitation through cross-linking of antigenic determinants. Immunodiffusion in agar gel revealed that M467 was recognizing more than one group of peptide determinants on the flagellins studied. Passive hemagglutination inhibition and a solid phase radioimmunoassay provided evidence that there were differences in binding avidities between M467 and the various Salmonella flagellins studied. It was concluded that M467 is binding more than one specific group of antigenic peptide determinants on flagellin molecules. Flagellin from four of the seven species of Salmonella studied were deficient in one or more of these determinants.

Animals↗
PubMed1979

Impairment of Jones-Mote hypersensitivity and specific antibody response against depolymerized flagellin in lepromatous leprosy.

Cutaneous hypersensitivity and antibody-producing capacity were assessed in patients with lepromatous leprosy with defective immunity, by immunizing them with monomeric flagellin from Salmonella adelaide. Results were compared with those of controls, matched for age and sex, derived from similar socioeconomic stratum, but without any defect of the immunological system. In contrast to the normal individuals, who showed Jones-Mote type of hypersensitivity, no lepromatous patient could mount any 'delayed-in-time' cutaneous hypersensivivity reaction against an intradermal challenge of monomeric flagellin. However, when immunized through the subcutaneous route, both groups could produce adequate amounts of specific serum antibody. In addition to this unique split tolerance found in all lepromatous patients, some patients showed low levels of 'natural' IgM antibody, reduced formation of specific antibody when immunized through the subcutaneous route, and incomplete maturation of IgG class of anti-flagellin antibody. When immunized by the intradermal route, however, production of both anti-flagellin antibody and maturation of IgG antibody was significantly inhibited in normal adults but not in lepromatous patients. Thus, contrary to the earlier concept of hyperactivity of the humoral immune apparatus in lepromatous leprosy, the present study detected B-cell hypofunction in some patients.

Adult↗
PubMed1978

[Molecular parameters of sensitization of tannin-treated erythrocytes with Salmonella d-flagellin].

A study was made of the molecular parameters of sensitization of tannin-treated sheep red blood cells with salmonella d-flagellin, depending on the protein dose, exposure, and temperature of the medium; the role played in this process by physical and chemical mechanisms was ascertained, the latter proving to determine the hemosensitization level. Senstivity of the erythrocytic diagnostic agent to the agglutinating action of the specific serum was maximal with a stable binding by a single red blood cell of about 200 000 flagellin molecules, although the limit of stable binding constituted 340 000 molecules per one red blood cell. On condition of saturation limit of the erythrocyte flagellin should block not over half of the cell surface. The temperature elevation during the diagnostic agent preparation is advantageous in technological respect, since it leads to reduction of flagellin expenditure and increases the sensitivity of the passive hemagglutination tests.

Animals↗
PubMed1975

Integration, at hag or elsewhere, of H2 (phase-2 flagellin) genes transduced from Salmonella to Escherichia coli.

A fla mutant of E. coli K12 was given fla+ and H1-i by phage P1kc cotransduction from S. typhimurium, then made Fla- by transduction of ah1 from S. typhimurium. Motile clones expressing a Salmonella phase-2 antigen, e,n,x or 1,2, were obtained from the K12 i ah1 (therefore Fla-) line by P1kc transduction of flagellin-specifying genes, H2-e,n,x or H2-1,2, from Salmonella donors. Of eighteen such transductants sixteen failed to show phase variation, and on transduction back to Salmonella each structural gene for a phase-2 flagellin (or at least for its antigenically determinant part) now behaved as an allele of H1, presumably in consequence of incorporation in the hag region of the K12 recipient, in place of H1-i ah1. The e,n,x- and 1,2-specifying genes were shown to have been integrated in the K12 chromosome without the linked H1-repressor gene or the adjacent vh2 gene (controlling rate of phase-variation) and they responded to the repressing activity of an H2 allele elsewhere in the cell, in this respect resembling H1 alleles of Salmonella or hag alleles of E. coli. Two K12 e,n,x transductants had flagellin-specifying genes which when transduced back to Salmonella were integrated at H2; they are inferred to have resulted from integration of H2-e,n,x in the K12 chromosome elsewhere than the hag region. These two clones showed phase variation, between a Fla+ phase, with antigen e,n,x, and a Fla- phase (with e,n,x determinant in the nonactive state and the determinant of antigen i inactivated by ah1). The two integrated e,n,x genes when in the "active" state retained the ability to repress expression of exogenote H1 alleles, which indicates that the closely linked H1-repressor gene also was integrated. One of the two exceptional transductants derived its e,n,x gene from a Salmonella donor with the linked vh2- gene, which in Salmonella almost entirely prevents change of phase, and transduction of this e,n,x gene back to Salmonella recipients proved that vh2- had been incorporated into the E. coli chromosome along with the e,n,x determinant and the H1-repressor gene. The high frequency of change of phase (Fla+ in equilibrium Fla-) in the K12 e,n,x vh2- transductant concerned suggests that vh2- fails to prevent frequent change of state of the phase-determined part of H2 when vh2- and H2 are incorporated in the E. coli chromosome.

Bacterial Proteins↗
PubMed1978

[Immunogenic properties of various forms of flagellin].

The authors obtained and studied immunologically flagellae of S. typhi and flagellins (monomeric and polymeric) obtained from them. Immunological activity of flagellin was the greater the higher the molecular weight of the preparation (the polymer molecules were the largest and the most immunogenic). There were revealed differences between the preparations according to the dependence of the immunizing effect on the dose and the capacity to induce synthesis of 19S- and 7S-antibodies. A sharp change of the immunogenicity of flagellin preparation following gel-filtration through sepharose 2B was demonstrated.

Animals↗
PubMed1976

Amino acid sequence of flagellin of Bacillus subtilis 168. III. Tryptic peptides, N-bromosuccinimide peptides, and the complete amino acid sequence.

Of the 28 expected tryptic peptides from Bacillus subtilis 168 flagellin, 24 were isolated and sequenced. Several overlapping tryptic peptides were also characterized. Studies were also performed on two peptides of 142 and 162 residues isolated after cleavage of the flagellin molecule at the single tyrosine residue (residue 142) with N-bromosuccinimide. These studies together with the previous data on the cyanogen bromide peptides and the tryptic peptides from maleylated flagellin permitted the complete amino acid sequence to be established: (see article). The primary structure reveals no obvious regularities or major repetitions of homologous sequences. Hydrophobic residues are distributed randomly in the amino acid sequence. However, the distribution of charged residues is strikingly asymmetric. The NH2-terminal region (residues 1 to 101) possesses a net charge of 6 plus, the middle of the molecule (residues 102 to 203), a net charge of 9 minus, and the COOH terminal region (residues 204 to 304), a net charge of 4 minus.

Amino Acid Sequence↗
PubMed1977

Classification of Bacillus subtilis flagellins.

Purified flagellins derived from 16 strains of Bacillus subtilis were classified into at least five distinct groups on the basis of their reaction with antiflagellar filament antibody and antiflagellin antibody. This classification was in good accord with that derived independently on the basis of amino acid analyses of the flagellins. Flagellar antigenicity appears to provide a useful typological character in classifying B. subtilis strains.

Amino Acids↗
PubMed1977

Caulobacter flagellins.

Flagellins from 17 Caulobacter strains were partially purified. Their molecular weights and immunological properties indicated that a two-component flagellin system is common among Caulobacters.

Bacteria↗
PubMed1978

Factors influencing the secondary antibody response to flagellin in man.

The secondary antibody response to 5.0 microgram flagellin was studied by haemagglutination in 132 healthy or convalescent subjects given a primary challenge with 5.0 microgram flagellin from 1 to 44 months previously. The peak titre, expressed as total antibody, occurred at 2 weeks and was mainly immunoglobulin (Ig)G. The magnitude of the titre of total antibody was influenced predominantly by that of total antibody in the primary response (P less than 0.001), the interval between primary and secondary responses (P less than 0.005) and the subjects' age (P less than 0.05) and sex (P less than 0.08). Together these accounted for 23% of the variability observed in the secondary response, with total antibody titre in the primary response accounting for 11% of the variability. The titre of IgG antibody was likewise influenced by these four variables, but the influence of age or sex on IgG antibody was not statistically significant. In human vaccination programmes, choice of the appropriate interval between primary and booster inoculations could increase prophylactic effectiveness and, if two inoculations were to prove as effective as three, there would be reduced work and increased public acceptance. Moreover, the demonstrable capacity for responsiveness of aged and debilitated persons should encourage the wider use of appropriate prophylactic immunization in these groups.

Adolescent↗
PubMed1976

Amino acid sequence of flagellin of Bacillus subtilis 168. I. Cyanogen bromide peptides.

Sequence studies are presented on the cyanogen bromide peptides derived from the flagellin of Bacillus subtilis 168. Eight unique CNBr peptides, ranging in length from 4 to 113 residues, were isolated in pure state. These peptides accounted for the amino acid composition of flagellin. The NH2-terminal methionyl residue of the protein reacted only partially with CNBr. Partial cleavage was observed at a Met-Glu bond (residues 22 to 23) of the protein.

Amino Acid Sequence↗
PubMed1978

Cell-mediated immunity to bacterial flagellin as assessed by leucocyte adherence inhibition.

Leucocyte adherence inhibition (LAI) was used to detect cell-mediated immunity of mice to Salmonella adelaide polymeric flagellin and its monomeric derivative. In the direct LAI technique, antigen inhibited the in vitro adherence to glass of peritoneal cells (PC) from antigen-primed mice which were capable of exhibiting in vivo delayed hypersensitivity reactions to the same antigen. In the indirect technique, primed PC exposed to antigen in vitro released a soluble factor, which inhibited the adherence of normal PC. Production of the factor was prevented by prior treatment of primed PC with anti-theta serum, indicating the participation of T-lymphocytes. The LAI reaction could be blocked by serum from mice which had been re-injected with antigen 72 h after a priming injection. Features of the production and biological properties of serum blocking activity suggest that it may be attributed to antigen-antibody complexes.

Animals↗
PubMed1976

Antigen-initiated B-lymphocyte differentiation. VIII. Sedimentation velocity and buoyant density characterization of virgin antibody-forming cell progenitors in the adoptive immune response of unprimed CBA mice to 4-hydroxy-3-iodo-5-nitrophenylacetic acid-polymerized bacterial flagellin antigen.

The characteristics of antibody-forming cell (AFC) progenitors lacking previous contact with specific antigen (virgin AFC progenitors) has been studied using sedimentation velocity and buoyant density separation for the investigation of physically distinct B-cell subpopulations. Functional characterization of isolated subsets was made using a quantitative adoptive immune assay for the IgM AFC progenitors responding to the antigen 4-hydroxy-3-iodo-5-nitrophenylacetic acid conjugated polymerized bacterial flagellin. Extensive heterogeneity is present among B lymphocytes, only some subpopulations of which exhibit AFC progenitor function. In the spleen of adult conventional CBA mice, atypically fast sedimenting cells of low buoyant density are active, while typical small B lymphocytes do not appear to be progenitors of IgM AFC. Spleen of adult specific pathogen-free (SPF), germfree, and athymic nude mice give similar results, although a minor population of typical slowly sedimenting dense cells are active in the latter two sources. Adult conventional bone marrow cells are as physically and functionally heterogeneous as splenic B cells, and although a significant proportion of AFC progenitor activity is found among dense, slowly sedimenting cells, most of the activity is among low density, faster sedimenting cells. In contrast to this situation in adult animals, where most of the unprimed AFC progenitors are large, atypical B cells, the spleens of neonatal mice provide a site where virgin AFC progenitors with the physical properties of typical small B lymphocytes are found. While being present in conventional and SPF neonatal spleens, these virgin cells are predominant in 7-day-old germfree mouse spleen. These findings suggest that the newborn virgin B cell is a typical small lymphocyte. However, few cells of this type are found in the adult animal. The unprimed AFC-progenitor population in the adult consists of large, fast sedimenting, low buoyant density, adherent cells, the physical properties of which are characteristic of activated B lymphocytes. It is suggested that these atypical cells are derived from the small newborn virgin B cell by the nonspecific effects of environmental antigenic stimuli.

Age Factors↗
PubMed1979

Regulation of expression of the flagellin gene (hag) in Escherichia coli K-12: analysis of hag-lac gene fusions.

Previous studies have defined 28 genes necessary for the synthesis of the flagellar apparatus of Escherichia coli K-12. This study analyzed the influence of the flagellar genes on the expression of the hag gene (structural gene for flagellin). To this end, a hag::Mu d(Apr lac) mutant which had the lac genes fused to the promoter of the hag gene was constructed. This allowed the measurement of hag gene expression by detection of beta-galactosidase activity. The following observations were made. (i) The hag gene was expressed constitutively in Fla+ cells. (ii) hag gene expression was positively regulated by flaA, FLAB, flaC, flaD, flaE, flaG, flaH, flaI, flaK, flaL, flaM, flaN, flaO, flaP, flaQ, flaR, flaV, flaW, flaX, flaY, flaZ, flbA, and flbB genes.hag-lac expression was not observed in strains with these fla mutations. (iii) The hag gene was expressed in mutants with flaS, flaT, flaU, and flbC defects. Therefore, these genes were not involved in regulation of hag gene transcription.

Bacterial Proteins↗