Dataset Information

Complete nucleotide sequence of the bacteriophage K1F tail gene encoding endo-N-acylneuraminidase (endo-N) and comparison to an endo-N homolog in bacteriophage PK1E.

ABSTRACT: Endo-N-acylneuraminidase (endo-N) is a phage-encoded depolymerase that degrades the alpha (2-8)-linked polysialic acid chains of K1 serotypes of Escherichia coli and vertebrate neural cell adhesion molecules. We have determined the DNA sequence of the bacteriophage K1F tail protein structural gene, which codes for a polypeptide of 920 residues. Purification of the tail protein yields a 102-kDa species upon denaturing gel electrophoresis and detection by Western immunoblot analysis. An identical polypeptide was detected by Western blot analysis of K1F virions. Peptide sequencing confirmed that the open reading frame determined by nucleotide sequencing encodes endo-N. Immunoelectron microscopy with neutralizing antibodies raised against the depolymerase confirmed that endo-N is a component of the K1F tail apparatus. Antibodies in the serum cross-reacted with endo-N from another K1-specific phage, PK1E, demonstrating the presence of shared epitopes. Homology between K1F and PK1E endo-N was confirmed by Southern, Northern (RNA), and Western blot analyses. The endo-N amino-terminal domain is homologous to the amino termini of phage T7 and T3 tail proteins, indicating by analogy that this domain functions in attachment of endo-N to the K1F virion's head. A central domain of 495 residues has weak similarity to sea urchin aryl sulfatase, suggesting that this region may contain the endo-N catalytic site. Failure to detect homology between the PK1E homolog and the carboxy-terminal domain of K1F endo-N is consistent with the central domain's involvement in binding and catalysis of polysialic acid. These results provide the initial molecular and genetic description of polysialic acid depolymerase, which has so far been detected only in K1-specific phage.

SUBMITTER: Petter JG

PROVIDER: S-EPMC204875 | biostudies-other | 1993 Jul

REPOSITORIES: biostudies-other

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Complete nucleotide sequence of the bacteriophage K1F tail gene encoding endo-N-acylneuraminidase (endo-N) and comparison to an endo-N homolog in bacteriophage PK1E.

Petter J G JG Vimr E R ER

Journal of bacteriology 19930701 14

Endo-N-acylneuraminidase (endo-N) is a phage-encoded depolymerase that degrades the alpha (2-8)-linked polysialic acid chains of K1 serotypes of Escherichia coli and vertebrate neural cell adhesion molecules. We have determined the DNA sequence of the bacteriophage K1F tail protein structural gene, which codes for a polypeptide of 920 residues. Purification of the tail protein yields a 102-kDa species upon denaturing gel electrophoresis and detection by Western immunoblot analysis. An identical ...[more]

PMID: 8331067

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Project description:The merozoite surface protein-1 (MSP1) is a prime candidate for an asexual blood stage vaccine against malaria. However, polymorphism in this antigen could compromise the vaccine's efficacy. Although the extent of sequence variation in MSP1 has been analyzed from various Plasmodium species, little is known about structural organization and diversity of this locus in Plasmodium malariae (PmMSP1). Herein, we have shown that PmMSP1 contained five conserved and four variable blocks based on analysis of the complete coding sequences. Variable blocks were characterized by short insertion and deletion variants (block II), polymorphic nonrepeat sequences (block IV), complex repeat structure with size variation (block VI) and degenerate octapeptide repeats (block VIII). Like other malarial MSP1s, evidences of intragenic recombination have been found in PmMSP1. The rate of nonsynonymous nucleotide substitutions significantly exceeded that of synonymous nucleotide substitutions in block IV, suggesting positive selection in this region. Codon-based analysis of deviation from neutrality has identified a codon under purifying selection located in close proximity to the homologous region of the 38 kDa/42 kDa cleavage site of P. falciparum MSP1. A number of predicted linear B-cell epitopes were identified across both conserved and variable blocks of the protein. However, polymorphism in repeat-containing blocks resulted in alteration of the predicted linear B-cell epitope scores across variants. Although a number of predicted HLA-class II-binding peptides were identified in PmMSP1, all variants of block IV seemed not to be recognized by common HLA-class II alleles among Thai population, suggesting that diversity in this positive selection region could probably affect host immune recognition. The data on structural diversity in PmMSP1 could be useful for further studies such as vaccine development and strain characterization of this neglected malaria parasite.

Dataset Information

Complete nucleotide sequence of the bacteriophage K1F tail gene encoding endo-N-acylneuraminidase (endo-N) and comparison to an endo-N homolog in bacteriophage PK1E.

Publications

Complete nucleotide sequence of the bacteriophage K1F tail gene encoding endo-N-acylneuraminidase (endo-N) and comparison to an endo-N homolog in bacteriophage PK1E.

Similar Datasets

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