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Is antiquitin a mitochondrial Enzyme?
Author(s) -
Wong Judy WeiYan,
Chan ChiLung,
Tang WaiKwan,
Cheng Christopher HonKi,
Fong WingPing
Publication year - 2009
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.22381
Subject(s) - cell fractionation , mitochondrion , western blot , subcellular localization , cytosol , biology , biochemistry , microbiology and biotechnology , transfection , hek 293 cells , mitochondrial matrix , enzyme , gene
Antiquitin is an aldehyde dehydrogenase involved in the catabolism of lysine. Mutations of antiquitin have been linked with the disease pyridoxine‐dependent seizures. While it is well established that lysine metabolism takes place in the mitochondrial matrix, evidence for the mitochondrial localization of antiquitin has been lacking. In the present study, the subcellular localization of antiquitin was investigated using human embryonic kidney HEK293 cells. Three different approaches were used. First, confocal microscopic analysis was carried out on cells transiently transfected with fusion constructs containing enhanced green fluorescent protein with different lengths of antiquitin based on the different potential start codons of translation. Second, immunofluorescence staining was used to detect the localization of antiquitin directly in the cells. Third, subcellular fractionation was carried out and the individual fraction was analyzed for the presence of antiquitin by Western blot and flow cytometric analyses. All the results showed that antiquitin was present not only in the cytosol but also in the mitochondria. J. Cell. Biochem. 109: 74–81, 2010. © 2009 Wiley‐Liss, Inc.