Western blotting analysis of the beta-hexosaminidase alpha- and beta-subunits in cultured fibroblasts from cases of various forms of GM2 gangliosidosis.

Abstract

OBJECTIVES:

The GM2 gangliosidoses are a group of genetic disorders caused by the accumulation of ganglioside GM2 in neuronal cells. We examined the alpha- and beta-subunits of beta-hexosaminidases by a non-radioisotopes detecting system to evaluate whether it was a useful method for understanding of the pathophysiologies of GM2 gangliosidoses.

MATERIALS AND METHODS:

We investigated the alpha- and beta-subunits of beta-hexosaminidases in cultured fibroblasts from cases of various forms of GM2 gangliosidosis by means of Western blotting and a chemiluminescence detection system.

RESULTS:

In a patient with infantile Tay-Sachs disease [HEXA genotype, Int5-SA(g-1-->t)/Int5-SA(g-1-->t)], the mature alpha-subunit was undetectable. In a patient with infantile Sandhoff disease (HEXB genotype, C534Y/C534Y), the mature beta-subunit was deficient. However, a small amount of the mature beta-subunit was detected in a patient with adult Sandhoff disease (HEXB genotype, R505Q(+I207V)/R505Q(+I207V)), which may have resulted in the residual enzyme activity and mild clinical course. Normal amounts of alpha- and beta-subunits were detected in a patient with GM2 activator deficiency.

CONCLUSION:

This method is easy and sensitive for detecting target proteins, and is useful for clarification of the pathophysiologies of GM2 gangliosidoses.

• Authors: K Utsumi, A Tsuji, R Kase, A Tanaka, T Tanaka, E Uyama, T Ozawa, H Sakuraba, Y Komaba, M Kawabe, Y Iino, Y Katayama
• Journal: Acta neurologica Scandinavica (Acta Neurol Scand) Vol. 105 Issue 6 Pg. 427-30 (Jun 2002) ISSN: 0001-6314 [Print] Denmark
• PMID: 12027830 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies
• Hexosaminidase A
• Hexosaminidase B
• beta-N-Acetylhexosaminidases

Topics

• Adult
• Antibodies
• Blotting, Western
• Cells, Cultured
• Female
• Fibroblasts (chemistry, cytology)
• Gangliosidoses, GM2 (metabolism, pathology)
• Hexosaminidase A
• Hexosaminidase B
• Humans
• Infant
• Luminescent Measurements
• Male
• Sandhoff Disease (metabolism, pathology)
• Skin (cytology)
• Tay-Sachs Disease (metabolism, pathology)
• beta-N-Acetylhexosaminidases (analysis, immunology)