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Cloning and sequence analysis of a cDNA clone coding for the mouse GM2 activator protein
G Bellachioma1, J L Stirling, A Orlacchio
1Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Universita di Perugia, Italy.
The Biochemical Journal
|August 15, 1993
Summary
Researchers isolated the mouse GM2 activator protein (GM2AP) cDNA, revealing a 579 bp coding sequence and a long 3' untranslated region. Mouse and human GM2AP sequences share significant identity, suggesting conserved function.
Area of Science:
- Molecular Biology
- Biochemistry
Background:
- GM2 activator protein (GM2AP) is crucial for the enzymatic hydrolysis of GM2 gangliosides.
- Understanding GM2AP's structure and function is vital for lysosomal storage disease research.
Purpose of the Study:
- To isolate and characterize the mouse GM2 activator protein (GM2AP) cDNA.
- To compare the mouse GM2AP sequence with its human counterpart.
Main Methods:
- cDNA library screening using a human GM2AP probe.
- Northern blot analysis of mouse macrophage RNA.
- Sequence alignment and hydropathicity plotting of deduced amino acid sequences.
Main Results:
- A 1.1 kb mouse GM2AP cDNA was isolated, featuring a 579 bp open reading frame and a ~1.7 kb 3' untranslated region.
- Mouse and human GM2AP sequences exhibit 68% overall amino acid identity, with 75% identity in the C-terminal region.
- Both sequences show similar hydropathicity profiles, indicating conserved structural features.
- A single N-glycosylation site was identified in mouse GM2AP at Asn151, differing in location from the human protein.
Conclusions:
- The mouse GM2AP gene structure, including a long 3' UTR, is conserved with the human gene.
- Significant sequence and structural similarities suggest conserved functional roles for GM2AP in both species.
- Differences in N-glycosylation site location may indicate species-specific post-translational modifications.