Processive phosphorylation of ERK MAP kinase in mammalian cells.

• Aoki, Kazuhiro

• Yamada, Masashi

• Kunida, Katsuyuki

• Yasuda, Shuhei

• Matsuda, Michiyuki

Abstract

The mitogen-activated protein (MAP) kinase pathway is comprised of a three-tiered kinase cascade. The distributive kinetic mechanism of two-site MAP kinase phosphorylation inherently generates a nonlinear switch-like response. However, a linear graded response of MAP kinase has also been observed in mammalian cells, and its molecular mechanism remains unclear. To dissect these input-output behaviors, we quantitatively measured the kinetic parameters involved in the MEK (MAPK/ERK kinase)-ERK MAP kinase signaling module in HeLa cells. Using a numerical analysis based on experimentally determined parameters, we predicted in silico and validated in vivo that ERK is processively phosphorylated in HeLa cells. Finally, we identified molecular crowding as a critical factor that converts distributive phosphorylation into processive phosphorylation. We proposed the term quasi-processive phosphorylation to describe this mode of ERK phosphorylation that is operated under the physiological condition of molecular crowding. The generality of this phenomenon may provide a new paradigm for a diverse set of biochemical reactions including multiple posttranslational modifications.

Journal

• Proceedings of the National Academy of Sciences of the United States of America

  Proceedings of the National Academy of Sciences of the United States of America 108 (31), 12675-12680, 2011-07-18

  National Academy of Sciences

Keywords

• simulation
• phos-tag
• crowder
• quantitative parameter
• EGF

Details

• CRID: 1050282676917642368
• NII Article ID: 120003204017
• NII Book ID: AA10808769
• ISSN: 00278424
• HANDLE: 2433/143060
• Text Lang: en
• Article Type: journal article
• Data Source:
  • IRDB
  • CiNii Articles