Molecular characterization of two myoglobins of Paragonimus westermani.
Myoglobins (Mbs), globin proteins, are present in high concentrations in trematodes. In Paragonimus westermani, 2 cDNAs were found to encode Mbs. The first clone, Pwmyo1, codes a total of 149 amino acids with a calculated mass of 16.6 kDa. The second, Pwmyo2, encodes a 146-amino acid protein with a calculated mass of 16.2 kDa. The predicted secondary structures showed the presence of 8 helices, which is the basic characteristic of Mbs. Sequence alignment revealed a high homology with the other trematode Mbs. The 2 clones contained the characteristic tyrosyl residues at helical positions B10 and distal E7, which are substitutions that have been previously shown to contribute to the high oxygen affinity of Mbs. Polyclonal antibodies against the recombinant Mbs were raised with no cross-reactivity observed. Immunolocalization revealed the proteins to be distributed generally throughout the parenchymal tissues, but absent from the tegument and reproductive organs. The cell mass of the eggs of the worm stained positive to Pwmyo2 but not Pwmyo1, suggesting the stage-specific expression of these Mbs.
de Guzman
Jefferson V
JV
Department of Parasitology, Kyungpook National University School of Medicine, Daegu 700-422, Korea.
Yu
Hak-Sun
HS
Jeong
Hae-Jin
HJ
Hong
Yeon-Chul
YC
Kim
Jin
J
Kong
Hyun-Hee
HH
Chung
Dong-Il
DI
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis / Q000276:immunology; D015153:Blotting, Western; D018076:DNA, Complementary / Q000737:chemistry*; D018509:DNA, Helminth / Q000737:chemistry*; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D015723:Gene Library; D007150:Immunohistochemistry; D007526:Isoelectric Point; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009211:Myoglobin / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D016366:Open Reading Frames / Q000235:genetics; D048492:Paragonimus westermani / Q000737:chemistry* / Q000145:classification / Q000235:genetics; D010802:Phylogeny; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015183:Restriction Mapping; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D000909:Antibodies, Helminth; D018076:DNA, Complementary; D018509:DNA, Helminth; D009211:Myoglobin; D011994:Recombinant Proteins