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Isolation and expression of a full-length cDNA encoding the human GM2 activator protein.
- Source :
-
Biochemical and biophysical research communications [Biochem Biophys Res Commun] 1991 Jun 28; Vol. 177 (3), pp. 1217-23. - Publication Year :
- 1991
-
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.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Cell Line
Cloning, Molecular
DNA isolation & purification
G(M2) Activator Protein
G(M2) Ganglioside metabolism
HeLa Cells physiology
Humans
Kinetics
Molecular Sequence Data
Oligonucleotide Probes
Polymerase Chain Reaction methods
Protein Sorting Signals genetics
Protein Sorting Signals metabolism
Proteins metabolism
Recombinant Proteins metabolism
Transfection
DNA genetics
Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0006-291X
- Volume :
- 177
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Biochemical and biophysical research communications
- Publication Type :
- Academic Journal
- Accession number :
- 2059210
- Full Text :
- https://doi.org/10.1016/0006-291x(91)90671-s