Isolation and expression of a full-length cDNA encoding the human GM2 activator protein.
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Xie B, McInnes B, Neote K, Lamhonwah AM, Mahuran D
Isolation and expression of a full-length cDNA encoding the human GM2 activator protein.
Biochem Biophys Res Commun. 1991 Jun 28;177(3):1217-23.
- PubMed ID
- 2059210 [ View in PubMed]
- Abstract
We report the construction of a cDNA clone encoding a functional GM2-activator protein. The sequence of the complete 5' end of the coding region was determined by direct nucleotide sequencing of a fragment generated by multiple RACE PCR procedures from Hela cell cDNA. Specific oligonucleotides were synthesized from these data which allowed us to produce a PCR fragment that contained the complete coding sequence of the protein. This was then cloned into a mammalian expression vector. The ability of purified hexosaminidase A (beta-N-acetylhexosaminidase, EC 3.2.1.52) to hydrolyse labeled GM2 ganglioside was enhanced 10-fold more by the addition in the assay mix of lysate from transfected COS-1 cells than by the addition of identical amounts of lysate from mock transfected cells. Direct sequencing of PCR fragments from two sources also identified three polymorphisms.