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Donald P Mcmanus - One of the best experts on this subject based on the ideXlab platform.

  • update on Paramyosin in parasitic worms
    Parasitology International, 2005
    Co-Authors: Geoffrey N Gobert, Donald P Mcmanus
    Abstract:

    Paramyosin was first identified as a structural component of invertebrate muscle. Analysis of crude, native, adult schistosome worm preparations identified a highly immunogenic protein which was later identified as Paramyosin. Early vaccination/challenge studies with native Paramyosin produced encouraging levels of protective efficacy against schistosomes, which led to the question as to how a sub-tegumental (muscular) protein could provide a target for vaccine-mediated immunological attack. Immunolocalisation studies of schistosomes confirmed the presence of Paramyosin within the post-acetabular glands of cercariae and on the tegumental surface of lung schistosomula. Here we present an update on the more recent research on Paramyosin in parasitic worms that has focused primarily in two directions: (i) further testing of the vaccine potency of Paramyosin against schistosomes and other parasitic worms; and (ii) characterisation of the protein at the molecular and biochemical levels.

  • the vaccine efficacy of native Paramyosin sj 97 against chinese schistosoma japonicum
    International Journal for Parasitology, 1998
    Co-Authors: Donald P Mcmanus, Guangchen Song, Yuxian Xu, Joanna M Wong
    Abstract:

    Abstract One of the promising anti-schistosome vaccine candidates currently under investigation is Paramyosin, a 97-kDa myofibrillar protein located in the muscles and tegument of schistosome worms. Here we describe the results of two vaccination\challenge experiments undertaken in mice using native Paramyosin isolated from adult worms of a Chinese strain of Schistosoma japonicum . In both sets of experiments, a relatively low but consistent and significant reduction in worm burden was evident in mice vaccinated subcutaneously with S. japonicum Paramyosin and Freunds adjuvant. In contrast, intraperitoneal vaccination of mice with Chinese strain S. japonicum Paramyosin without adjuvant did not result in any reduction in worm numbers when compared with a saline control group. These data contrast with the impressive protection figures reported by another group who used a similar intraperitoneal vaccination protocol with native Paramyosin extracted from Philippine strain S. japonicum .

  • engineering and expression of a full length cdna encoding schistosoma japonicum Paramyosin purification of the recombinant protein and its recognition by infected patient sera
    Acta Tropica, 1997
    Co-Authors: Bernd H Kalinna, Marion M Becker, Donald P Mcmanus
    Abstract:

    Abstract A cDNA encoding the complete open reading frame of the Schistosoma japonicum Paramyosin has been constructed and cloned. The 2600 bp cDNA was engineered by PCR using a N-terminally truncated Paramyosin clone (pmy25) as template and a 57-mer primer that introduced the eight missing amino acids and matched the 5′ sequence of pmy25 in conjunction with a pmy specific reverse primer. After cloning and expression, the recombinant protein was purified by affinity chromatography under non-denaturing conditions and was shown to have a molecular mass of 99 kDa which is equivalent to the expected size of the full length recombinant fusion protein, comprising the 97 kDa Paramyosin plus an additional 2 kDa for the N-terminal fusion peptide incorporating the six histidine residues required for purification. In Western blot assays it reacted specifically with anti-Paramyosin antibodies in sera from vaccinated animals and patients with Asian schistosomiasis. The engineering of the full-length cDNA encoding Schistosoma japonicum Paramyosin, its bacterial expression and purification will facilitate future studies aimed at determining its efficacy as an anti-schistosomiasis vaccine.

  • schistosoma japonicum immunolocalization of Paramyosin during development
    Parasitology, 1997
    Co-Authors: Geoffrey N Gobert, Deborah J Stenzel, Malcolm K Jones, D E Allen, Donald P Mcmanus
    Abstract:

    This paper describes the localization of Paramyosin immunoreactivity in Schistosoma japonicum and represents the first comparative immunolocalization study among schistosome adult, cercariae and lung schistosomula by electron microscopy. A polyclonal antibody was utilized to immunolabel Paramyosin or Paramyosin-like proteins. Paramyosin was localized within the muscle layer of all 3 developmental stages. Furthermore, Paramyosin was localized within granules of the post-acetabular glands of cercariae, and within the tegument matrix and surface of lung schistosomules. Adults and cercariae did not display any detectable Paramyosin on the surface or within the tegument. The possible functions of Paramyosin within S. japonicum and the relevance of these findings in relation to the reported protective properties of Paramyosin as an anti-schistosome vaccine target molecule are discussed.

  • schistosoma japonicum heterogeneity in Paramyosin genes
    Acta Tropica, 1995
    Co-Authors: William C Hooker, Donald P Mcmanus, Wen Yang, Marion M Becker, G J Waine, Bernd H Kalinna, Karen A Nimmo, Paul J Brindley
    Abstract:

    Abstract Paramyosin is an integral muscle protein found in many invertebrates including schistosomes, and is considered an important candidate vaccine antigen in schistosomiasis. The characterisation of natural molecular variation in vaccine antigens including Paramyosin is important because strain-specific vaccination may be necessary against schistosomiasis japonica. We have isolated partial cDNAs encoding Paramyosin from an adult, Chinese strain Schistosoma japonicum gene library. Two of these cDNAs (B6 and Y6) encode the same region of Paramyosin but their nucleotide sequences differ at eight positions and their deduced amino acid sequences differ by an arginine/cysteine substitution, demonstrating intrastrain variation in Paramyosin. Southern blot analysis of genomic DNA from the Chinese and Philippine strains of S. japonicum demonstrated strain-related RFLPs at the Paramyosin locus, and suggested that more than one copy of the Paramyosin gene was present in the S. japonicum genome. PCR-based RFLP analysis which exploited restriction site differences between B6 and Y6 showed that Paramyosin genotype B6 was much more common in schistosome populations and verified the existence of introns in the Paramyosin gene(s) of S. japonicum .

Juan Pedro Laclette - One of the best experts on this subject based on the ideXlab platform.

  • gene structure of taenia solium Paramyosin
    Parasitology Research, 2003
    Co-Authors: Laura Vargasparada, Juan Pedro Laclette
    Abstract:

    Paramyosin is a muscle protein that probably plays a role in the survival of the larval stage of Taenia solium during its prolonged host-parasite relationship. Here we describe the structure of the gene coding for the Paramyosin of T. solium. The characterization of two clones obtained from a genomic library showed that the complete gene of Paramyosin contains 13 introns delimited by conventional eukaryotic splice signals. Comparison with the Paramyosin genes of Drosophila melanogaster and Caenorhabditis elegans showed a lack of conservation of the exon/intron organization in contrast to other muscle genes. No evidence of alternative splicing sites were found, excluding the possibility that T. solium expresses a mini-Paramyosin like D. melanogaster.

  • human t and b cell epitope mapping of taenia solium Paramyosin
    Parasite Immunology, 2001
    Co-Authors: Jose Vazqueztalavera, Carlos F Solis, Edith Medinaescutia, Zoila Morales Lopez, Jefferson V Proano, Dolores Correa, Juan Pedro Laclette
    Abstract:

    Taenia solium Paramyosin is an immunodominant antigen in human and porcine cysticercosis that has shown promise as a vaccine candidate against schistosomiasis and some filariasis. There are few studies to identify the immunologically relevant regions of Paramyosin. In this work, we characterize the humoral and cellular response of neurocysticercotic patients against T. solium Paramyosin. Western blots using different recombinant fragments of T. solium Paramyosin, showed that the sera from neurocysticercotic patients were strongly reactive against the carboxyl end region, with poor recognition of the central and amino regions. In contrast, the cellular immune response of patients did not show preferential recognition of any region of Paramyosin.

  • Paramyosin inhibits complement c1
    Journal of Immunology, 1992
    Co-Authors: Juan Pedro Laclette, Charles B Shoemaker, D Richter, Lourdes Arcos, N Pante, C Cohen, D Bing, Anne Nicholsonweller
    Abstract:

    We report here the results of studies showing that inhibition of C is a property of several invertebrate Paramyosins. Paramyosins from Taenia solium, Schistosoma mansoni, and the mussel Mytilus edulis bind polymeric collagen and can be isolated from crude extracts of tissues by collagen affinity. These Paramyosins inhibit C1 function whether the C1 is isolated or present in C2-deficient serum. Because T. solium Paramyosin was the best inhibitor, we concentrated further studies on this molecule. T. solium Paramyosin binds purified C1q in solution with a dose/response similar to C1r2S2. Further studies of the C1-Paramyosin interaction indicate that: 1) C4 is not activated, 2) C4b2a decay is not affected, and 3) there is no effect on the efficiency of C3-9, as provided in EDTA-chelated guinea pig serum, in lysing SRBC. Thus, Paramyosin inhibition is directed at the initiation of the classical pathway. The results suggest that Paramyosins of helminthic parasites may have a role as modulators of the host immune response through C inhibition at C1.

  • Paramyosin is the schistosoma mansoni trematoda homologue of antigen b from taenia solium cestoda
    Molecular and Biochemical Parasitology, 1991
    Co-Authors: Juan Pedro Laclette, Lourdes Arcos, Abraham Landa, Kaethe Willms, Alvin E Davis, Charles B Shoemaker
    Abstract:

    Abstract Antigen B, a major antigen of the cestode parasite Taenia solium, has been purified and a portion of amino acid sequence obtained. Paramyosin of the trematode parasite Schistosoma mansoni, an immunogenic protein that has shown promise as a vaccine candidate, has several biochemical and immunological properties in common with antigen B. A full-length cDNA clone of S. mansoni Paramyosin has been obtained and the predicted translation product contains a sequence that is highly homologous to the sequence obtained for antigen B. The predicted amino acid composition and isolectric point of Paramyosin are nearly identical to those established for antigen B. Recombinant S. mansoni Paramyosin, expressed in Escherichia coli as a fusion protein with β-galactosidase, was recognized by antisera against T. solium antigen B. We conclude from these results that S. mansoni Paramyosin and T. solium antigen B are hormologous proteins. Since S. mansoni Paramyosin is thought to be a muscle protein and T. solium antigen B a secreted glycoprotein with anti-complement activity, this conclusion raises some interesting questions regarding the role of this class of proteins in the host-parasite relationship.

Kazuo Shiomi - One of the best experts on this subject based on the ideXlab platform.

  • Paramyosin from the disc abalone haliotis discus discus
    Journal of Food Biochemistry, 2014
    Co-Authors: Midori Suzuki, Keiko Shimizu, Yukihiro Kobayashi, Shoichiro Ishizaki, Kazuo Shiomi
    Abstract:

    Besides tropomyosin that represents a major allergen in mollusks as well as in crustaceans, a 100-kDa allergen was recently found in the disc abalone Haliotis discus discus and identified as Paramyosin, an invertebrate-specific myofibrillar protein. In this study, the amino acid sequence (860 residues) of disc abalone Paramyosin was elucidated by cDNA cloning. As high as 70% amino acid sequence homology was recognized between disc abalone and Mediterranean mussel Paramyosins, supporting the previously suggested immunoglobulin E (IgE) cross-reactivity between both Paramyosins. Disc abalone Paramyosin was expressed in Escherichia coli as a HisGln (HQ)-tagged protein. As analyzed by enzyme-linked immunosorbent assay, the recombinant preparation was judged to have almost the same IgE-binding ability as its natural counterpart. Practical Applications This study showed that sufficient amounts of the recombinant disc abalone Paramyosin can be obtained in a relatively short time whenever needed. The recombinant disc abalone Paramyosin is comparable in IgE-binding ability to the natural counterpart and hence could be used as an alternative antigen of the natural counterpart for molecular studies and diagnosis of mollusk allergy.

  • Paramyosin of the disc abalone haliotis discus discus identification as a new allergen and cross reactivity with tropomyosin
    Food Chemistry, 2011
    Co-Authors: Midori Suzuki, Yukihiro Kobayashi, Yumiko Hiraki, Hiroki Nakata, Kazuo Shiomi
    Abstract:

    Abstract Tropomyosin, a 35–38 kDa myofibrillar protein, represents a major allergen in molluscs, as well as in crustaceans. Besides tropomyosin, a 100 kDa allergen was newly detected in the disc abalone Haliotis discus discus. The 100 kDa allergen was purified from the muscle of the disc abalone by salting-out and hydroxyapatite HPLC and identified as Paramyosin based on the determined amino acid sequences of the peptide fragments produced by lysylendopeptidase digestion. Based on analysis by fluorescence ELISA, as many as 16 of the 18 patient sera tested, reacted to the disc abalone tropomyosin. The same patient sera also reacted to the disc abalone Paramyosin, although rather less potently than to tropomyosin, suggesting that Paramyosin is a major allergen. Immunoblotting data showed that IgE-reactive Paramyosin is distributed in some species of molluscs other than the disc abalone. Interestingly, cross-reactivity between Paramyosin and tropomyosin was demonstrated by inhibition immunoblotting and inhibition ELISA.

Gary J Weil - One of the best experts on this subject based on the ideXlab platform.

  • immune responses to brugia malayi Paramyosin in rodents after dna vaccination
    Vaccine, 1999
    Co-Authors: Benwen Li, Shaorong Zhang, Kurt C Curtis, Gary J Weil
    Abstract:

    Immunization with recombinant Brugia malayi Paramyosin protein (BM5) induces partial immunity to this filarial nematode in jirds. The present study examined the effects of intramuscular immunization with plasmid DNA that encodes BM5. DNA-immunized mice produced strong antibody and cell-mediated responses to Paramyosin. The protective activity of DNA vaccination with BM5 was tested in jirds. Vaccinated jirds produced strong antibody responses to Paramyosin, but adult worm recoveries after challenge were not decreased in vaccinated animals relative to controls. These studies show that DNA vaccination can induce immune responses to filarial antigens in rodents. Further efforts will be needed to achieve the goal of inducing protective immunity to filariasis with this promising new technology.

  • vaccination with recombinant filarial Paramyosin induces partial immunity to brugia malayi infection in jirds
    Journal of Immunology, 1993
    Co-Authors: Benwen Li, Ramaswamy Chandrashekar, Gary J Weil
    Abstract:

    Vaccination with irradiated infective larvae induces partial protective immunity to infection with the filarial nematode Brugia malayi in jirds. Prior studies have shown that such immunization stimulates a much stronger antibody response to recombinant and native filarial Paramyosin than that seen after normal infection. To determine whether vaccination with recombinant Paramyosin could induce protective immunity to larval challenge, jirds were immunized with either B. malayi Paramyosin and maltose binding protein (BM5-MBP) (fusion protein of B. malayi Paramyosin and maltose-binding protein), MBP alone, or recombinant Dirofilaria immitis Paramyosin. Animals were challenged with 100 infective larvae s.c. 8 wk after the second immunization. Necropsies were performed 16 wk after challenge. Vaccination with BM5-MBP induced significant protective immunity; adult worm recoveries, worm lengths, and blood microfilaria counts were reduced in the BM5-MBP group relative to the MBP control group. The reductions in adult worm recoveries (43%) and female worm lengths (10%) were statistically significant (p

Margarita Cervera - One of the best experts on this subject based on the ideXlab platform.

  • Paramyosin and MiniParamyosin
    Nature’s Versatile Engine: Insect Flight Muscle Inside and Out, 2020
    Co-Authors: Margarita Cervera, Juan J Arredondo, Raquel Marco Ferreres
    Abstract:

    In Drosophila, Paramyosin and miniParamyosin are structural components of thick filaments that have a similar structure to the myosin heavy chain rod tail. Both proteins are rod-like molecules with a high α-helical content in the long central domains, and exist as dimers. While miniParamyosin is mainly located in the M line and at both ends of the thick filaments in Drosophila indirect flight muscles (IFM), Paramyosin is present all along the thick filaments. The relative amounts of myosin, Paramyosin and miniParamyosin vary in the distinct muscles, reflecting the differences in the organization of their thick filaments. Moreover, as for other contractile proteins, the phosphorylation of these two proteins is involved in the acquisition of the capacity to fly. Thus, miniParamyosin has a possible role in the sequential transition of nonfunctional to functional muscle, in general, while the Paramyosin transition is more specifically related to the functional onset of IFM.

  • drosophila Paramyosin miniParamyosin gene products show a large diversity in quantity localization and isoform pattern a possible role in muscle maturation and function
    Journal of Cell Biology, 1996
    Co-Authors: Miguel Maroto, Belinda Bullard, Roberto Marco, Juan J Arredondo, David Goulding, Margarita Cervera
    Abstract:

    The Drosophila Paramyosin/miniParamyosin gene expresses two products of different molecular weight transcriptionally regulated from two different promoters. Distinct muscle types also have different relative amounts of myosin, Paramyosin, and miniParamyosin, reflecting differences in the organization of their thick filaments. Immunofluorescence and EM data indicate that miniParamyosin is mainly located in the M line and at both ends of the thick filaments in Drosophila indirect flight muscles, while Paramyosin is present all along the thick filaments. In the tergal depressor of the trochanter muscle, both proteins are distributed all along the A band. In contrast, in the waterbug, Lethocerus, both Paramyosin and miniParamyosin are distributed along the length of the indirect flight and leg muscle thick filaments. Two-dimensional and one-dimensional Western blot analyses have revealed that miniParamyosin has several isoforms, focusing over a very wide pH range, all of which are phosphorylated in vivo. The changes in isoform patterns of miniParamyosin and Paramyosin indicate a direct or indirect involvement of these proteins in muscle function and flight. This wide spectrum of potential regulatory characteristics underlines the key importance of Paramyosin/miniParamyosin and its complex isoform pattern in the organization of the invertebrate thick filament.

  • Analysis of the Paramyosin/MiniParamyosin Gene MINIParamyosin IS AN INDEPENDENTLY TRANSCRIBED, DISTINCT Paramyosin ISOFORM, WIDELY DISTRIBUTED IN INVERTEBRATES
    Journal of Biological Chemistry, 1995
    Co-Authors: Miguel Maroto, Roberto Marco, Juan J Arredondo, Marta Roman, Margarita Cervera
    Abstract:

    Abstract MiniParamyosin, a distinct Drosophila melanogaster Paramyosin isoform of 60 kDa, is shown here to be encoded by the same gene as Paramyosin. The gene, located at 66D14, spans over 12.8 kilobases (kb) and is organized into 10 exons, 9 of which code for the Paramyosin transcripts. An exon, located between exons 7 and 8, codes for the 5′-end of the miniParamyosin, and the two proteins share the two last exons of the gene. Mapping of the 5′-ends of these transcripts indicates that the Paramyosin and miniParamyosin mRNAs arise from two overlapping transcriptional units; the miniParamyosin transcription initiation site is located inside a Paramyosin intron, 8 kb downstream of the one used for Paramyosin transcription. The existence of two different promoters and the conserved and nonconserved features of their sequences suggest a very complex regulation of these two muscle proteins. In fact, while Paramyosin is expressed at two distinct stages of development as most other Drosophila muscle proteins, miniParamyosin appears late in development, being present only in the adult musculature. The absence of exon 1B, the specific exon of miniParamyosin, in the nematode Caenorhabditis elegans, as well as additional lines of evidence support the lack of miniParamyosin in this particular organism. However, it is present in most invertebrate species examined, including different arthropod, annelid, mollusc, and echinoderm species.

  • analysis of the Paramyosin miniParamyosin gene miniParamyosin is an independently transcribed distinct Paramyosin isoform widely distributed in invertebrates
    Journal of Biological Chemistry, 1995
    Co-Authors: Miguel Maroto, Roberto Marco, Juan J Arredondo, Marta Roman, Margarita Cervera
    Abstract:

    Abstract MiniParamyosin, a distinct Drosophila melanogaster Paramyosin isoform of 60 kDa, is shown here to be encoded by the same gene as Paramyosin. The gene, located at 66D14, spans over 12.8 kilobases (kb) and is organized into 10 exons, 9 of which code for the Paramyosin transcripts. An exon, located between exons 7 and 8, codes for the 5′-end of the miniParamyosin, and the two proteins share the two last exons of the gene. Mapping of the 5′-ends of these transcripts indicates that the Paramyosin and miniParamyosin mRNAs arise from two overlapping transcriptional units; the miniParamyosin transcription initiation site is located inside a Paramyosin intron, 8 kb downstream of the one used for Paramyosin transcription. The existence of two different promoters and the conserved and nonconserved features of their sequences suggest a very complex regulation of these two muscle proteins. In fact, while Paramyosin is expressed at two distinct stages of development as most other Drosophila muscle proteins, miniParamyosin appears late in development, being present only in the adult musculature. The absence of exon 1B, the specific exon of miniParamyosin, in the nematode Caenorhabditis elegans, as well as additional lines of evidence support the lack of miniParamyosin in this particular organism. However, it is present in most invertebrate species examined, including different arthropod, annelid, mollusc, and echinoderm species.

  • identification and characterization of drosophila melanogaster Paramyosin
    Journal of Molecular Biology, 1991
    Co-Authors: Javier Vinos, Alberto Domingo, Roberto Marco, Margarita Cervera
    Abstract:

    Abstract Paramyosin, a major structural component of thick filaments in invertebrates has been isolated, purified and characterized from whole adult Drosophila melanogaster extracts and a specific polyclonal antibody against it has been prepared. Paramyosin has been identified on the basis of several criteria, including molecular weight, α-helicity, species distribution, capability of fiber formation in vitro and sequence. We have used the immunopurified polyclonal antibody to isolate eight clones from a λgtl1 expression library of Drosophila 1 to 22 h embryo cDNA. The largest clone (pJV9) has been sequenced and encodes the coiled-coil region of D. melanogaster Paramyosin that is 47% identical to Caenorhabditis elegans Paramyosin. Indirect immunofluorescence in semi-thin sections of adult flies show fluorescence mainly in tubular muscle. Freshly prepared tubular myofibrils decorated with the immunoabsorbed antibody show the A region in the sarcomere as the specific localization of Paramyosin. The amount of Paramyosin in tubular synchronous muscles of insects appears to be five times higher than in fibrillar insect muscles. There are at least three Paramyosin isoforms as shown by isoelectrofocusing separation. The more acidic and less abundant form is phosphorylated as shown by 32P in vivo labeling experiments in adult flies. The developmental pattern of expression of Drosophila Paramyosin is presented. This mesoderm-specific protein, immunologically undetectable during gastrulation and early phases of germ band formation, progressively increases during organogenesis to the adult stage. Interestingly, it is also expressed as a major maternal product in the insoluble cytoskeletal fraction of the mature oocyte.