Sequence of a full-length cDNA for rat lung beta-galactoside-binding protein: primary and secondary structure of the lectin.
A full-length cDNA for rat lung beta-galactoside lectin (subunit Mr approximately 14,000, lectin 14K) was cloned and the nucleotide sequence determined. The deduced amino acid sequence agrees with the amino acid composition and direct amino acid sequence analysis of purified rat lung lectin peptides. We found that the amino-terminal alanine is blocked with an acetyl group. Comparison of the amino acid sequence with other proteins shows a high degree of homology only with other vertebrate lectin sequences, supporting the suggestion that these lectins may constitute a unique class of vertebrate proteins. The amino acid composition and sequence of lectin peptides, the sequence of lectin cDNA, and isoelectric focusing of purified lectin indicate that rat lung lectin 14K is composed predominantly of a single protein. In addition, rat uterus lectin 14K was found to be the same protein as that present in lung. We characterized the secondary and tertiary structure of rat lung lectin 14K by circular dichroism, by analytical ultracentrifugation, and by computer analysis of its primary structure. Results of these experiments suggest that lectin 14K is primarily a hydrophilic protein with an asymmetric, elongated structure consisting of approximately equal amounts of alpha helix, beta sheet, beta turn, and random coil. We found that Cys-2 and Cys-130 react most rapidly with iodoacetamide; one or both of these residues may be primarily responsible for the thiol requirement of lectin activity.
Clerch
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Calvin and Flavia Oak Asthma Research and Treatment Facility, Department of Medicine, Miami, Florida.
Whitney
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P
Hass
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Brew
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Miller
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Werner
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R
Massaro
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D005260:Female; D005696:Galactosidases / Q000378:metabolism; D037161:Galectins; D006388:Hemagglutinins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D051381:Rats; D011919:Rats, Inbred Strains; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014599:Uterus / Q000378:metabolism
D037161:Galectins; D006388:Hemagglutinins; D004247:DNA; D005696:Galactosidases