Novel immunoreactive glycoprotein orthologs of Ehrlichia spp.

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We previously identified a strongly immunoreactive 43 kDa protein (p43) of Ehrlichia canis. As an immunodiagnostic antigen, the p43 had a 96% accuracy as compared with IFA and provided species-specific diagnosis of E. canis infections. Further investigation has revealed that the E. canis p43 represents the N-terminal portion of the largest immunoreactive protein described in Ehrlichia spp. with a predicted molecular mass of 153 kDa. Analysis of the recombinant N-terminal region (p43) of the p153 by protein gel electrophoresis demonstrated a larger than predicted molecular mass (∼30%), and presence of carbohydrate glycans, indicating that the p153 is a glycoprotein. A BLASTn search was performed on the E. chaffeensis genome sequence (95%), and the gene encoding the p153 ortholog was identified in E. chaffeensis. The E. canis p153 (4,263 bp) and E. chaffeensis p156 (4,389 bp) genes had similar chromosomal locations, downstream of the homologous (∼87%) deoxyguanosine-triphosphate triphosphohydrolase genes, and homologous (∼90%) intergenic sequences preceding the open reading frames. Nucleic acid sequence homology (52%) observed between the glycoprotein genes supported previous findings with regard to genetic divergence of the p43 gene fragment, and the p153 and p156 proteins had amino acid similarity of 32%. A native E. canis protein with a molecular mass of 200 kDa reacted with antisera produced against the N-terminal region (p43) of the p153, suggesting that the native protein was posttranslationally modified. Similarly, recombinant constructs of E. chaffeensis p156 migrated larger than predicted (∼200 kDa), and carbohydrate was detected on the recombinant proteins. The chromosomal location, amino acid homology, and biophysical properties support the conclusion that the p153 and p156 glycoproteins (designated gp200s) are species-specific immunoreactive orthologs.

Original languageEnglish (US)
Pages (from-to)678-684
Number of pages7
JournalAnnals of the New York Academy of Sciences
Volume990
StatePublished - 2003

Fingerprint

Ehrlichia
Ehrlichia canis
Glycoproteins
Genes
Molecular mass
Proteins
Nucleic Acid Sequence Homology
Carbohydrates
Amino Acids
Nucleic acid sequences
Intergenic DNA
Gene encoding
Recombinant Proteins
Open Reading Frames
Polysaccharides
Electrophoresis
Immune Sera
Gels
Protein
Genome

Keywords

  • Anaplasma
  • Bacterial glycoproteins
  • Ehrlichia canis
  • Ehrlichia chaffeensis

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Novel immunoreactive glycoprotein orthologs of Ehrlichia spp. / McBride, Jere; Comer, Jason; Walker, David.

In: Annals of the New York Academy of Sciences, Vol. 990, 2003, p. 678-684.

Research output: Contribution to journalArticle

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title = "Novel immunoreactive glycoprotein orthologs of Ehrlichia spp.",
abstract = "We previously identified a strongly immunoreactive 43 kDa protein (p43) of Ehrlichia canis. As an immunodiagnostic antigen, the p43 had a 96{\%} accuracy as compared with IFA and provided species-specific diagnosis of E. canis infections. Further investigation has revealed that the E. canis p43 represents the N-terminal portion of the largest immunoreactive protein described in Ehrlichia spp. with a predicted molecular mass of 153 kDa. Analysis of the recombinant N-terminal region (p43) of the p153 by protein gel electrophoresis demonstrated a larger than predicted molecular mass (∼30{\%}), and presence of carbohydrate glycans, indicating that the p153 is a glycoprotein. A BLASTn search was performed on the E. chaffeensis genome sequence (95{\%}), and the gene encoding the p153 ortholog was identified in E. chaffeensis. The E. canis p153 (4,263 bp) and E. chaffeensis p156 (4,389 bp) genes had similar chromosomal locations, downstream of the homologous (∼87{\%}) deoxyguanosine-triphosphate triphosphohydrolase genes, and homologous (∼90{\%}) intergenic sequences preceding the open reading frames. Nucleic acid sequence homology (52{\%}) observed between the glycoprotein genes supported previous findings with regard to genetic divergence of the p43 gene fragment, and the p153 and p156 proteins had amino acid similarity of 32{\%}. A native E. canis protein with a molecular mass of 200 kDa reacted with antisera produced against the N-terminal region (p43) of the p153, suggesting that the native protein was posttranslationally modified. Similarly, recombinant constructs of E. chaffeensis p156 migrated larger than predicted (∼200 kDa), and carbohydrate was detected on the recombinant proteins. The chromosomal location, amino acid homology, and biophysical properties support the conclusion that the p153 and p156 glycoproteins (designated gp200s) are species-specific immunoreactive orthologs.",
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N2 - We previously identified a strongly immunoreactive 43 kDa protein (p43) of Ehrlichia canis. As an immunodiagnostic antigen, the p43 had a 96% accuracy as compared with IFA and provided species-specific diagnosis of E. canis infections. Further investigation has revealed that the E. canis p43 represents the N-terminal portion of the largest immunoreactive protein described in Ehrlichia spp. with a predicted molecular mass of 153 kDa. Analysis of the recombinant N-terminal region (p43) of the p153 by protein gel electrophoresis demonstrated a larger than predicted molecular mass (∼30%), and presence of carbohydrate glycans, indicating that the p153 is a glycoprotein. A BLASTn search was performed on the E. chaffeensis genome sequence (95%), and the gene encoding the p153 ortholog was identified in E. chaffeensis. The E. canis p153 (4,263 bp) and E. chaffeensis p156 (4,389 bp) genes had similar chromosomal locations, downstream of the homologous (∼87%) deoxyguanosine-triphosphate triphosphohydrolase genes, and homologous (∼90%) intergenic sequences preceding the open reading frames. Nucleic acid sequence homology (52%) observed between the glycoprotein genes supported previous findings with regard to genetic divergence of the p43 gene fragment, and the p153 and p156 proteins had amino acid similarity of 32%. A native E. canis protein with a molecular mass of 200 kDa reacted with antisera produced against the N-terminal region (p43) of the p153, suggesting that the native protein was posttranslationally modified. Similarly, recombinant constructs of E. chaffeensis p156 migrated larger than predicted (∼200 kDa), and carbohydrate was detected on the recombinant proteins. The chromosomal location, amino acid homology, and biophysical properties support the conclusion that the p153 and p156 glycoproteins (designated gp200s) are species-specific immunoreactive orthologs.

AB - We previously identified a strongly immunoreactive 43 kDa protein (p43) of Ehrlichia canis. As an immunodiagnostic antigen, the p43 had a 96% accuracy as compared with IFA and provided species-specific diagnosis of E. canis infections. Further investigation has revealed that the E. canis p43 represents the N-terminal portion of the largest immunoreactive protein described in Ehrlichia spp. with a predicted molecular mass of 153 kDa. Analysis of the recombinant N-terminal region (p43) of the p153 by protein gel electrophoresis demonstrated a larger than predicted molecular mass (∼30%), and presence of carbohydrate glycans, indicating that the p153 is a glycoprotein. A BLASTn search was performed on the E. chaffeensis genome sequence (95%), and the gene encoding the p153 ortholog was identified in E. chaffeensis. The E. canis p153 (4,263 bp) and E. chaffeensis p156 (4,389 bp) genes had similar chromosomal locations, downstream of the homologous (∼87%) deoxyguanosine-triphosphate triphosphohydrolase genes, and homologous (∼90%) intergenic sequences preceding the open reading frames. Nucleic acid sequence homology (52%) observed between the glycoprotein genes supported previous findings with regard to genetic divergence of the p43 gene fragment, and the p153 and p156 proteins had amino acid similarity of 32%. A native E. canis protein with a molecular mass of 200 kDa reacted with antisera produced against the N-terminal region (p43) of the p153, suggesting that the native protein was posttranslationally modified. Similarly, recombinant constructs of E. chaffeensis p156 migrated larger than predicted (∼200 kDa), and carbohydrate was detected on the recombinant proteins. The chromosomal location, amino acid homology, and biophysical properties support the conclusion that the p153 and p156 glycoproteins (designated gp200s) are species-specific immunoreactive orthologs.

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