Acetaldehyde substoichiometrically inhibits bovine neurotubulin polymerization.
Acetaldehyde is known to form covalent adducts with tubulin and to inhibit microtubule formation. Available evidence indicates that lysine residues are prominently involved in adduct formation. Previous work has shown that lysines on tubulin can be divided into two general classes based upon their reactivity toward acetaldehyde; those of normal reactivity ("bulk" lysines) and a highly reactive lysine (HRL) located on the alpha-polypeptide subunit. We took advantage of the fact that the HRL is unreactive when tubulin is in the microtubule form to differentiate the effects of bulk from HRL adducts on tubulin polymerization. Under conditions where both bulk lysines and HRL formed adducts, 0.2 mol acetaldehyde/mol tubulin caused complete inhibition of polymerization. When we modified bulk lysines, but not HRL, tubulin polymerized essentially normally. Finally, when we first blocked bulk lysines on microtubules (HRL unreactive) using unlabeled acetaldehyde and then measured the amount of [14C]acetaldehyde adduct formed with tubulin after depolymerization (HRL reactive), 0.08 mol acetaldehyde/mol tubulin resulted in completely impaired polymerization. These data show that microtubule formation is very sensitive to even small mole fractions of acetaldehyde-modified tubulin (especially with HRL) and further suggest that small amounts of acetaldehyde adduct could be damaging to cytoskeleton function in the cell.
Smith
S L
SL
Liver Study Unit, Veterans Administration Medical Center, Omaha, Nebraska 68105.
Jennett
R B
RB
Sorrell
M F
MF
Tuma
D J
DJ
D000079:Acetaldehyde / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D008239:Lysine / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects*; D011108:Polymers; D050257:Tubulin Modulators
D011108:Polymers; D050257:Tubulin Modulators; D000079:Acetaldehyde; D008239:Lysine