Details

Autor(en) / Beteiligte

• Riera, Thomas V.
• Wigle, Tim J.
• Gureasko, Jodi
• Boriack-Sjodin, P. Ann
• Copeland, Robert A.

Titel

Abstract 2144: Kinetic mechanism of the lysine methyltransferase SMYD3 using MAP3K2 protein substrate

Ist Teil von

• Cancer research (Chicago, Ill.), 2015-08, Vol.75 (15_Supplement), p.2144-2144

Erscheinungsjahr

2015

Quelle

EZB Free E-Journals

Beschreibungen/Notizen

• Abstract Dysregulation of DNA and histone methylation has been shown to play fundamental roles in carcinogenesis. Recently however, protein methylation in the cytoplasm has been linked to oncogenic Ras signaling. Mazur and colleagues demonstrated that SMYD3 (SET and MYND domain containing protein 3), a primarily cytoplasmic protein methyltransferase, can methylate MAP3K2 using S-adenosyl-L-methionine (SAM) as a cofactor (Mazur et al. 2014. Nature 510(7504):283-7). Methylation by SMYD3 preserves the active, phosphorylated state of MAP3K2 sustaining activation of the Ras/Raf/MEK/ERK/ signaling module and potentiating formation of Ras-driven cancers. Here, we characterized the kinetic mechanism of SMYD3 using MAP3K2 protein as a substrate. SMYD3 activity was measured with a radiometric assay using 3H-SAM. Initial velocity, product and dead-end inhibitor studies indicate that SMYD3 uses a random kinetic mechanism where both SAM and MAP3K2 can bind apo-SMYD3. This was supported by substrate and ligand binding assays using SPR. Together, these studies elucidate the kinetic mechanism of SMYD3 with MAP3K2 and provide a context for inhibitor development. Citation Format: Thomas V. Riera, Tim J. Wigle, Jodi Gureasko, P. Ann Boriack-Sjodin, Robert A. Copeland. Kinetic mechanism of the lysine methyltransferase SMYD3 using MAP3K2 protein substrate. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2144. doi:10.1158/1538-7445.AM2015-2144