Sequence analysis of the 47-kilodalton major integral membrane immunogen of Treponema pallidum

P. L. Hus, N. R. Chamberlain, K. Orth, C. R. Moomaw, L. Q. Zhang, Clive A. Slaughter, J. D. Radolf, S. Sell, M. V. Norgard

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The complete primary amino acid sequence for the 47-kilodalton (kDa) major integral membrane immunogen of Treponema pallidum subsp. pallidum was obtained by using a combined strategy of DNA sequencing (of the cloned gene in Escherichia coli) and N-terminal amino acid sequencing of the native (T. pallidum subsp. pallidum-derived) antigen. An open reading frame believed to encode the 47-kDa antigen comprised 367 amino acid codons, which gave rise to a calculated molecular weight for the corresponding antigen of 40,701. Of the 367 amino acids, 113 (31%) were sequenced by N-terminal amino acid sequencing of trypsin and hydroxylamine cleavage fragments of the native molecule isolated from T. pallidum subsp. pallidum; amino acid sequence data had a 100% correlation with that of the amino acid sequence predicted from DNA sequencing of the cloned gene in E. coli. Although no consensus sequences for the initiation of transcription or translation were readily identifiable immediately 5' to the putative methionine start codon, a 63-base-pair PstI fragment located 159 nucleotides upstream was required for expression of the 47-kDa antigen in E. coli. The 47-kDa antigen sequence did not reveal a typical leader sequence. The overall G+C content for the DNA corresponding to the structural gene was 53%. Hydrophilicity analysis identified at least one major hydrophilic domain of the protein near the N terminus of the molecule which potentially represents an immunodominant epitope. No repetitive primary sequence epitopes were found. The combined data provide the molecular basis for further structural and functional studies regarding the role of the antigen in the immunopathogenesis of treponemal disease.

Original languageEnglish (US)
Pages (from-to)196-203
Number of pages8
JournalInfection and Immunity
Volume57
Issue number1
StatePublished - Jan 1 1989
Externally publishedYes

Fingerprint

Treponema pallidum
Sequence Analysis
Antigens
Globus Pallidus
Membranes
Amino Acid Sequence
Protein Sequence Analysis
Escherichia coli
DNA Sequence Analysis
Genes
Amino Acids
Immunodominant Epitopes
Hydroxylamine
Initiator Codon
Nucleic Acid Repetitive Sequences
Consensus Sequence
Base Composition
Hydrophobic and Hydrophilic Interactions
Codon
Base Pairing

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

Cite this

Hus, P. L., Chamberlain, N. R., Orth, K., Moomaw, C. R., Zhang, L. Q., Slaughter, C. A., ... Norgard, M. V. (1989). Sequence analysis of the 47-kilodalton major integral membrane immunogen of Treponema pallidum. Infection and Immunity, 57(1), 196-203.

Sequence analysis of the 47-kilodalton major integral membrane immunogen of Treponema pallidum. / Hus, P. L.; Chamberlain, N. R.; Orth, K.; Moomaw, C. R.; Zhang, L. Q.; Slaughter, Clive A.; Radolf, J. D.; Sell, S.; Norgard, M. V.

In: Infection and Immunity, Vol. 57, No. 1, 01.01.1989, p. 196-203.

Research output: Contribution to journalArticle

Hus, PL, Chamberlain, NR, Orth, K, Moomaw, CR, Zhang, LQ, Slaughter, CA, Radolf, JD, Sell, S & Norgard, MV 1989, 'Sequence analysis of the 47-kilodalton major integral membrane immunogen of Treponema pallidum', Infection and Immunity, vol. 57, no. 1, pp. 196-203.
Hus PL, Chamberlain NR, Orth K, Moomaw CR, Zhang LQ, Slaughter CA et al. Sequence analysis of the 47-kilodalton major integral membrane immunogen of Treponema pallidum. Infection and Immunity. 1989 Jan 1;57(1):196-203.
Hus, P. L. ; Chamberlain, N. R. ; Orth, K. ; Moomaw, C. R. ; Zhang, L. Q. ; Slaughter, Clive A. ; Radolf, J. D. ; Sell, S. ; Norgard, M. V. / Sequence analysis of the 47-kilodalton major integral membrane immunogen of Treponema pallidum. In: Infection and Immunity. 1989 ; Vol. 57, No. 1. pp. 196-203.
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abstract = "The complete primary amino acid sequence for the 47-kilodalton (kDa) major integral membrane immunogen of Treponema pallidum subsp. pallidum was obtained by using a combined strategy of DNA sequencing (of the cloned gene in Escherichia coli) and N-terminal amino acid sequencing of the native (T. pallidum subsp. pallidum-derived) antigen. An open reading frame believed to encode the 47-kDa antigen comprised 367 amino acid codons, which gave rise to a calculated molecular weight for the corresponding antigen of 40,701. Of the 367 amino acids, 113 (31{\%}) were sequenced by N-terminal amino acid sequencing of trypsin and hydroxylamine cleavage fragments of the native molecule isolated from T. pallidum subsp. pallidum; amino acid sequence data had a 100{\%} correlation with that of the amino acid sequence predicted from DNA sequencing of the cloned gene in E. coli. Although no consensus sequences for the initiation of transcription or translation were readily identifiable immediately 5' to the putative methionine start codon, a 63-base-pair PstI fragment located 159 nucleotides upstream was required for expression of the 47-kDa antigen in E. coli. The 47-kDa antigen sequence did not reveal a typical leader sequence. The overall G+C content for the DNA corresponding to the structural gene was 53{\%}. Hydrophilicity analysis identified at least one major hydrophilic domain of the protein near the N terminus of the molecule which potentially represents an immunodominant epitope. No repetitive primary sequence epitopes were found. The combined data provide the molecular basis for further structural and functional studies regarding the role of the antigen in the immunopathogenesis of treponemal disease.",
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