title: Inactivation of GAPDH as one mechanism of acrolein toxicity.
abstract: We have recently reported that acrolein is more toxic than reactive oxygen species. Thus, the mechanism of cell toxicity by acrolein was studied using mouse mammary carcinoma FM3A cells. Acrolein-conjugated proteins were separated by gel electrophoresis with subsequent determination of their amino acid sequence, and it was found that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was one of the major acrolein-conjugated proteins in cells. Acrolein interacted with cysteine-150 at the active site of GAPDH, and also with cysteine-282. When cells were treated with 8 μM acrolein, the activity of acrolein-conjugated GAPDH was greatly reduced, and the ATP content in cells was thus significantly reduced. In addition, it was shown that acrolein-conjugated GAPDH translocated to the nucleus, and the level of acetylated GAPDH and the number of TUNEL positive cells was increased, indicating that cell death is enhanced by acrolein-conjugated GAPDH. Inhibition of cell growth by acrolein was partially reversed when the cDNA encoding GAPDH was transformed into cells. These results indicate that inactivation of GAPDH is one mechanism that underlies cell toxicity caused by acrolein.
authors.affiliation: Faculty of Pharmacy, Chiba Institute of Science, Choshi, Chiba, Japan.
authors.forename: Mizuho
authors.initials: M
authors.lastname: Nakamura
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Tomitori
authors.forename: Takehiro
authors.initials: T
authors.lastname: Suzuki
authors.forename: Akihiko
authors.initials: A
authors.lastname: Sakamoto
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Terui
authors.forename: Ryotaro
authors.initials: R
authors.lastname: Saiki
authors.forename: Naoshi
authors.initials: N
authors.lastname: Dohmae
authors.forename: Kazuei
authors.initials: K
authors.lastname: Igarashi
authors.forename: Keiko
authors.initials: K
authors.lastname: Kashiwagi
mesh_terms: D000171:Acrolein / Q000378:metabolism* / Q000633:toxicity*; D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D014170:Transformation, Genetic
chemical_list: D000171:Acrolein; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D003545:Cysteine