Characterization and immunolocalization of a Cryptosporidium protein containing repeated amino acid motifs.
The oocyst wall is one of the components that permits cryptosporidia both to survive in the environment and to retain infectivity. With the aim of identifying Cryptosporidium proteins specifically expressed at the oocyst stage, we screened lambda gt11 genomic libraries of Cryptosporidium parvum with both an oocyst antiserum and a specific genetic probe. We isolated, from distinct libraries, two overlapping clones containing an open reading frame encoding a 1,252-amino-acid polypeptide. The analysis of the deduced amino acid sequence revealed unusually high contents of cysteine, proline, and histidine. The sequence was also characterized by two distinct amino acid motifs, each repeated several times. The DNA sequences coding for the amino acid repeats showed a high frequency of synonymous mutations, a result suggesting that the repeated motifs may be functionally and/or structurally important to the parasite. Antisera and monoclonal antibodies developed against a recombinant polypeptide encompassing the first 786 amino acids revealed that the corresponding protein in C. parvum had an apparent molecular weight of 190,000. Moreover, confocal microscopy analysis with immunofluorescence indicated that the protein was localized on the oocyst wall as a uniform stain and within the oocyst itself as bright granules in close association with the residual body.
Ranucci
L
L
Istituto di Parassitologia, Università di Roma, La Sapienza, Italy.
Müller
H M
HM
La Rosa
G
G
Reckmann
I
I
Morales
M A
MA
Spano
F
F
Pozio
E
E
Crisanti
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D000918:Antibody Specificity; D001483:Base Sequence; D003458:Cryptosporidium / Q000737:chemistry* / Q000276:immunology; D004251:DNA Transposable Elements; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D017386:Sequence Homology, Amino Acid
D000913:Antibodies, Protozoan; D004251:DNA Transposable Elements; D015800:Protozoan Proteins; C081386:cpMM1 protein, Cryptosporidium parvum; C081387:cpRL3 protein, Cryptosporidium