Molecular cloning, characterization, and expression of a human 14-kDa lectin.
Full length cDNAs coding for a 14-kDa beta-galactoside binding lectin have been isolated from HL-60 cells and human placenta. Oligonucleotide probes based on a pentapeptide present in several partial sequences of homologous human lectins were used to screen a lambda GT10 HL-60 cDNA library. The HL-60 cDNA clones that were isolated were used to design a synthetic primer representing the 3'-untranslated region of the HL-60 lectin. This primer was then used to synthesize a lambda GT10 human placenta cDNA library, and restriction fragments of the HL-60 cDNA clones were used to screen the library. The cDNA clones for both HL-60 and placenta lectin had identical sequences with short 5'- and 3'-untranslated regions and coded for a 135-amino acid protein which lacks a hydrophobic signal peptide sequence. Biochemical data show that, despite the presence of a possible N-linked glycosylation site, the protein is not glycosylated. Northern and Southern blot analyses indicate that the 14-kDa lectin is encoded for by a single gene. The lectin cDNA was expressed in Escherichia coli and biologically active protein was purified from cell lysates by affinity chromatography.
Couraud
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Ideon Corporation, Redwood City, California 94063.
Casentini-Borocz
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D
Bringman
T S
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Griffith
J
J
McGrogan
M
M
Nedwin
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GE
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005260:Female; D037161:Galectins; D005796:Genes; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy
D037161:Galectins; D006388:Hemagglutinins; D004247:DNA