Molecular cloning of a human cDNA encoding β-1,4-galactosyltransferase with 37% identity to mammalian UDP-Gal:GlcNAc β-1,4-galactosyltransferase

DOI Web Site 被引用文献14件
  • Takeshi Sato
    Department of Biosignal Research, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo 173, Japan; and Department of Medical Chemistry, Vrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
  • Kiyoshi Furukawa
    Department of Biosignal Research, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo 173, Japan; and Department of Medical Chemistry, Vrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
  • Hans Bakker
    Department of Biosignal Research, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo 173, Japan; and Department of Medical Chemistry, Vrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
  • Dirk H. Van den Eijnden
    Department of Biosignal Research, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo 173, Japan; and Department of Medical Chemistry, Vrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands
  • Irma Van Die
    Department of Biosignal Research, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo 173, Japan; and Department of Medical Chemistry, Vrije Universiteit, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands

抄録

<jats:p> A cDNA encoding a β-1,4-galactosyltransferase named β-1,4-GalT II was cloned from a cDNA library of the human breast tumor cell line, MRK-nu-1. Initially, a 860-bp PCR fragment was obtained from MRK-nu-1 mRNA by 3′-rapid amplification of cDNA ends by using two nested degenerate oligonucleotide primers based on a highly conserved amino acid sequence found in the catalytic domain of mammalian β-1,4-galactosyltransferases and <jats:italic>Lymnaea stagnalis</jats:italic> β-1,4- <jats:italic>N</jats:italic> -acetylglucosaminyltransferase (β-1,4-GlcNAcT), both of which utilize the same sugar acceptor. This subsequently was used as a probe to isolate a 4.7-kb cDNA that contained an ORF of 1,164 bp predicting a polypeptide of 388 aa. Its deduced amino acid sequence shows an identity of 37% with that of the previously characterized human β-1,4-galactosyltransferase (referred to as β-1,4-GalT I) and of 28% with that of <jats:italic>L. stagnalis</jats:italic> β-1,4-GlcNAcT. Study of the properties of the β-1,4-GalT II fused to protein A expressed as a soluble form in COS-7 cells revealed that it is a genuine β-1,4-GalT but has no lactose synthetase activity in the presence of α-lactalbumin. Northern blot analysis of 24 human tissues showed that they all express the β-1,4-GalT II transcript, although the levels varied. These results indicate that human cells contain another β-1,4-GalT. </jats:p>

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