Cloning, expression, and chromosomal mapping of a human ganglioside sialidase

Tadashi Wada, Yuko Yoshikawa, Satoru Tokuyama, Masaaki Kuwabara, Hirotoshi Akita, Taeko Miyagi

Research output: Contribution to journalArticlepeer-review

119 Citations (Scopus)

Abstract

Here we report the cDNA sequence of a human ganglioside sialidase. The cDNA was isolated from a human brain cDNA library by screening with a 240 bp probe generated by polymerase chain reaction using primers based on the sequences of rat cytosolic and bovine membrane sialidases which we previously cloned. The 3.0 kb cDNA encodes an open reading frame of 436 amino acids containing a putative transmenbrane domain and an Arg-Ile-Pro and three Asp-box sequences characteristic of sialidases and showing overall 83% and 39% identities to the bovine and rat enzymes, respectively. Northern blot analysis revealed high expression in skeletal muscle and testis, but low level in kidney, placenta, lung, and digestive organs. Transient expression of the cDNA in COS-1 cells resulted in a 130-fold increase in sialidase activity compared to the control level, and the activity was found to be almost specific for gangliosides. Fluorescent in situ hybridization allowed the human sialidase gene localized to chromosome 11 at q 13.5.

Original languageEnglish
Pages (from-to)21-27
Number of pages7
JournalBiochemical and biophysical research communications
Volume261
Issue number1
DOIs
Publication statusPublished - 1999 Jul 22

Keywords

  • Chromosomal mapping
  • Gangliosides
  • Sialidase
  • cDNA cloning

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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