Molecular cloning, characterization, and expression of a human 14-kDa lectin

J Biol Chem. 1989 Jan 15;264(2):1310-6.

Abstract

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.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Cell Line
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular*
  • DNA / genetics
  • Female
  • Galectins
  • Genes
  • Hemagglutination Tests
  • Hemagglutinins / genetics*
  • Hemagglutinins / isolation & purification
  • Humans
  • Leukemia, Promyelocytic, Acute
  • Molecular Sequence Data
  • Molecular Weight
  • Placenta / metabolism
  • Pregnancy

Substances

  • Galectins
  • Hemagglutinins
  • DNA

Associated data

  • GENBANK/J04456