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In vitro binding of (14C)2,5-hexanedione to rat neuronal cytoskeletal proteins.
Lanning CL; Wilmarth KR; Abou-Donia MB
Neurochem Res 1994 Sep; 19(9):1165-1173
A study was conducted examining the binding of 2,5-hexanedione (110134) (2,5-HD) to neurofilaments and microtubule proteins in- vitro. Partially purified neurofilament and microtubule preparations were incubated with radiolabeled 2,5-HD and analyzed using sodium- dodecyl-sulfate polyacrylamide gel electrophoresis and immunoblotting. 2,5-HD was found to bind to cytoskeletal and microtubule proteins in a concentration and time dependent fashion. The highest amount of binding was seen to NFM followed by NFH and finally NFL. The binding of 2,5-HD to MAP-2 was five times that of its binding to tubulin. The rate of binding of 2,5-HD to MAP-2 was higher than its rate of binding to any other cytoskeletal protein. NFL and tubulin had the lowest rates of binding. High molecular weight materials with positive reactions to antibodies against NFH and NFM were seen in immunoblots of both control and treated samples. Analysis of spectral data of 2,5-HD in micellar and nonmicellar solutions over a 5 day period demonstrated increased absorbance as a function of time at multiple wavelengths in both solutions. The greatest relative absorbance difference between solutions was seen in the range of 460 nanometers where it was increased three fold in micellar solutions. The authors conclude that 2,5-HD binds to both neurofilaments and microtubules.
Neurotoxic-effects; Aliphatic-hydrocarbons; Toxic-effects; Protein-biochemistry; Subcellular-fractions; Neurotoxicity; In-vitro-studies; Author Keywords: Aliphatic hexacarbons; 2,5-hexanedione; axonopathy; cytoskeletal proteins
Pharmacology Duke University Department of Pharmacology Durham, N C 27710
Issue of Publication
Neurotoxic Disorders; Neurotoxic-effect
Duke University, Durham, North Carolina
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