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Western blotting analysis of the β‐hexosaminidase α‐ and β‐subunits in cultured fibroblasts from cases of various forms of GM2 gangliosidosis
Author(s) -
Utsumi K.,
Tsuji A.,
Kase R.,
Tanaka A.,
Tanaka T.,
Uyama E.,
Ozawa T.,
Sakuraba H.,
Komaba Y.,
Kawabe M.,
Iino Y.,
Katayama Y.
Publication year - 2002
Publication title -
acta neurologica scandinavica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.967
H-Index - 95
eISSN - 1600-0404
pISSN - 0001-6314
DOI - 10.1034/j.1600-0404.2002.01097.x
Subject(s) - sandhoff disease , gangliosidosis , hexosaminidase , tay sachs disease , hexa , biology , genotype , adenine phosphoribosyltransferase , microbiology and biotechnology , endocrinology , medicine , enzyme , biochemistry , disease , gene , purine
Objectives – The GM2 gangliosidoses are a group of genetic disorders caused by the accumulation of ganglioside GM2 in neuronal cells. We examined the α ‐ and β ‐subunits of β ‐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 α ‐ and β ‐subunits of β ‐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 α ‐subunit was undetectable. In a patient with infantile Sandhoff disease ( HEXB genotype, C534Y/C534Y), the mature β ‐subunit was deficient. However, a small amount of the mature β ‐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 α ‐ and β ‐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.