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R L Coppel

Publications and source records attributed to R L Coppel.

At least 163 records · Page 9Linked to original sources
PubMed1987

A cDNA clone expressing a rhoptry protein of Plasmodium falciparum.

Antibodies from immune humans were used to select a cDNA clone expressing an asexual blood stage antigen of Plasmodium falciparum. The expressed fused polypeptide was used as an affinity reagent to purify human antibodies specific for the corresponding parasite antigen. Western blotting and immunoelectronmicroscopy demonstrated that the antigen was a 105 kDa protein located in the rhoptries of merozoites. The cDNA encodes the carboxy terminus of the rhoptry antigen, a sequence rich both in charged and hydroxy amino acids.

Amino Acid Sequence↗
PubMed1987

Putative glycophorin-binding protein is secreted from schizonts of Plasmodium falciparum.

A cDNA clone expressing an antigen of Plasmodium falciparum, selected by screening an expression library cloned in Escherichia coli, encodes a portion of the protein identified as a glycophorin-binding protein [Kochan et al. (1986) Cell 44, 689-696]. Human antibodies affinity-purified on extracts from this clone were used to characterize the antigen by immunoblotting. This protein was present in all isolates tested, restricted to mature trophozoites and schizonts. It was abundant in culture supernatants at the time of merozoite release but present in minor amounts if at all in merozoites. The pattern of antigen distribution over schizont-infected cells observed by immunoelectron microscopy differed from that of the precursor of the major merozoite surface antigens in that most of the antigen appeared to be located over the erythrocyte cytoplasm without any obvious association with organelles. It thus appears unlikely that this antigen is present on the merozoite surface prior to schizont rupture.

Amino Acid Sequence↗
PubMed1987

Changes in repeat number, sequence, and reading frame in S-antigen genes of Plasmodium falciparum.

The S antigens from different isolates of Plasmodium falciparum exhibit extensive size, charge, and serological diversity. We show here that the S-antigen genes behave as multiple alleles of a single locus. The size heterogeneity results from different numbers, lengths, and/or sequences of tandem repeat units encoded within the S-antigen genes. Two genes studied here encode antigenically different S antigens but nevertheless have closely related tandem repeat sequences. We show that antigenic differences can arise because repeats are translated in different reading frames.

Alleles↗
PubMed1987

Sequence variation in S-antigen genes of Plasmodium falciparum.

S-antigens are soluble heat-stable antigens released into the circulation at the time of schizogony of Plasmodium falciparum. Many serologically distinct S-antigens exist and we have shown that this diversity results from repetitive sequences that vary in repeat number, length, sequence and/or reading frame among different S-antigens. We present here the complete sequence of the S-antigen of a Vietnamese isolate V1. The major repeat of 33 base-pairs can be considered to be derived by a deletion event from a 45 base-pair sequence that is located at the 3' repeat boundary and is related in sequence to all S-antigen repeats known so far. We also show that the non-repetitive coding region of the S-antigen gene of V1 is identical to that of K1 and only two amino acids different to that of NF7. In contrast, the sequences are considerably different to those of the FC27 and Wellcome isolates. We conclude that S-antigen genes are highly polymorphic in the repetitive regions but show more restricted diversity in non-repetitive regions.

Amino Acid Sequence↗
PubMed1986

Structure of the RESA gene of Plasmodium falciparum.

We have determined the nucleotide sequence of the gene encoding the ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum, an antigen that has been shown to confer protective immunity on monkeys. The sequence has enabled us to predict the structure of the RESA gene and the amino acid sequence of its protein product. The gene consists of two exons with a short intron located near the 5' end of the coding region. A hydrophobic amino acid segment predicted for the 3' end of exon 1 is consistent with the possibility that exon 1 encodes trafficking signal sequences. We show that restriction fragment length polymorphisms can be used to define two different alleles of RESA, represented by isolates FC27 and NF7, and compare the FC27 sequence with that of a long cDNA clone from NF7 described previously.

Amino Acid Sequence↗
PubMed1986

An asparagine-rich protein from blood stages of Plasmodium falciparum shares determinants with sporozoites.

We describe a cDNA clone derived from mRNA of asexual blood-stages of the malaria parasite Plasmodium falciparum. This clone, designated Ag319, expresses a P.falciparum antigen fused to beta-galactosidase in Escherichia coli. Human antibodies from Papua New Guinea were affinity-purified by adsorption to extracts of Ag319 immobilized on CNBr-Sepharose. The antibodies reacted predominantly with P. falciparum polypeptides of Mr 220,000 and 160,000, and a number of ill-defined lower molecular weight species. Antibodies reacted in indirect immunofluorescence with all asexual blood-stages although the antigen appeared to be most abundance in the schizont. Surprizingly the antibodies also reacted with sporozoites. The amino acid sequence predicted from the complete nucleotide sequence of this clone is remarkable because 40% of the residues are Asn, and so the antigen has been termed the Asparagine-Rich Protein (ARP). Like other P. falciparum antigens, ARP contains tandemly repetitive sequences, based on the tetrapeptide Asn-Asn-Asn-Met and we have confirmed that these represent natural epitopes by reaction of the corresponding synthetic peptides with human antibodies. Surprisingly, ARP is also rich in Asn outside the tandem repeats.

Amino Acid Sequence↗
PubMed1986

Affinity purification of human antibodies directed against cloned antigens of Plasmodium falciparum.

A technique has been developed for the affinity purification of antibodies recognizing cloned antigens of the malaria parasite Plasmodium falciparum expressed in bacteria. Adsorbents prepared by coupling bacterial lysates to Sepharose were used to isolate monospecific antibodies from human immune sera. Production of an abundant stable fused polypeptide by the bacteria was not a prerequisite for the success of this approach. Also the procedure permits the characterization of antigens which elicit the production of very low levels of antibodies. Affinity-purified human antibodies were used to characterized the corresponding P. falciparum antigens by immunoblotting and a number of antigens identified in this way illustrate some commonly observed features of P. falciparum antigens. Several of these antibody preparations recognized multiple bands in the electrophoretic patterns. Studies on a number of isolates of P. falciparum indicate that many antigens exhibit size polymorphisms. Production of some antigens was shown to be restricted to particular stages of the asexual blood cycle of the parasite while others appear to be specifically processed during the life cycle. Affinity-purified antibodies have also been used to locate antigens within the infected erythrocyte and to delineate subsets of antibodies recognizing different epitopes of a single antigen.

Animals↗
PubMed1986

Antigenic repeat structures in proteins of Plasmodium falciparum.

The majority of malaria antigens that have been cloned contain short sequence repeats which encode antigenic epitopes that are naturally immunogenic. Synthetic peptides have been used to show that natural antibody responses to a strain-specific Plasmodium falciparum S antigen are largely directed against epitopes encoded in an 11-amino acid sequence that is repeated approximately 100 times in the molecule. A 16-amino acid peptide conjugated to bovine serum albumin induced antibodies specific for the S antigen of the homologous isolate. Synthetic peptides have also been used to confirm the natural immunogenicity of epitopes encoded within two blocks of related repeats in the Ring-infected Erythrocyte Surface Antigen (RESA). A 16-amino acid peptide, comprising four repeats of the tetrameric sequence EENV, induced antibodies reactive with the native molecule. Detailed analyses of these anti-peptide antisera indicate that short sequence repeats express more than one epitope, some of which may cross-react with other repeat structures.

Amino Acid Sequence↗
PubMed1986

Variable antigen associated with the surface of erythrocytes infected with mature stages of Plasmodium falciparum.

Immune human sera were used to select a cDNA clone expressing an asexual blood-stage antigen of Plasmodium falciparum. Antibodies affinity-purified on extracts from this clone were used to characterize the antigen by immunoblotting and immunofluorescence. The antigen is present in mature-stage parasites as a high molecular weight protein of about 250 kDa and is apparently processed to smaller fragments in the merozoite. It varies in molecular weight and antibody reactivity in different isolates, and has been localized at the erythrocyte membrane by immunoelectronmicroscopy. Part of the protein is composed of exactly repeated hexapeptide units that constitute the strain-specific determinant. This molecule has similar characteristics to the strain-specific molecule believed to be responsible for cytoadherence.

Amino Acid Sequence↗
PubMed1986

A repetitive antigen of Plasmodium falciparum that is homologous to heat shock protein 70 of Drosophila melanogaster.

We describe an antigen of Plasmodium falciparum, defined by a cDNA clone designated Ag63. The antigen is an abundant, soluble cytoplasmic polypeptide of Mr 75,000 present in all stages of asexual development in the blood and in gametocytes, but not in sporozoites. The sequence of the cDNA clone revealed that, like many other antigens of P. falciparum, it contains tandemly repeated amino acid sequences, in this case Gly-Gly-Met-Pro. However, the rest of the sequence is 70% homologous at the amino acid level to the heat shock protein hsp70 of Drosophila melanogaster.

Amino Acid Sequence↗
PubMed1986

Prospects for a malaria vaccine.

Malaria infection is a worsening problem throughout the developing world where conventional methods of control and treatment are becoming ineffective. Recent discoveries using the tools of the new biology, monoclonal antibodies and gene cloning, have brought malaria vaccines to the brink of reality.

Animals↗
PubMed1986

Sorting large numbers of clones expressing Plasmodium falciparum antigens in Escherichia coli by differential antibody screening.

We describe an approach to classifying a large number of clones expressing Plasmodium falciparum antigens in Escherichia coli by virtue of their differing reactivities with 100 human anti-malarial sera. Individual sera exhibited marked differences in the patterns of reactivity with these clones. These patterns led to the identification of sets of clones, here termed "serological families", which were shown to encode distinct P. falciparum antigens. A serological family was found to be composed of non-identical clones derived from portions of the same antigen. Using this approach six new P. falciparum antigens were identified. One of these is described in detail and is a 102 X 10(3) Mr antigen, predominantly of schizonts. Sequencing studies on four cDNA clones encoding parts of this antigen revealed blocks of hydrophilic dipeptide and tripeptide repeats and so the antigen has been termed the acidic basic repeat antigen (ABRA).

Animals↗
PubMed1985

Sequence of a cDNA encoding a small polymorphic histidine- and alanine-rich protein from Plasmodium falciparum.

We describe the expression in Escherichia coli, isolation by immunological screening and complete nucleotide sequence of a cDNA clone from the malaria parasite Plasmodium falciparum. The deduced amino acid sequence contains separate blocks of repetitive hexapeptide and pentapeptide sequences and we have confirmed that these represent epitopes by reaction of the corresponding synthetic peptides with human antibodies. As the predicted size is Mr 21,000 and the overall composition is 30% His and 29% Ala, the polypeptide has been termed the small histidine-alanine rich protein (SHARP). This polypeptide is highly polymorphic in different P. falciparum isolates and cross reacts immunologically with a distinct gene product of P. falciparum. Although it is related to the Histidine Rich Protein (HRP) of P. lophurae by virtue of its high His content, it shows no obvious sequence relationship to the HRP outside the repeats.

Amino Acid Sequence↗