Next Article in Journal
A Fully Sensorized Cooperative Robotic System for Surgical Interventions
Next Article in Special Issue
Synthesis of Bioactive Microcapsules Using a Microfluidic Device
Previous Article in Journal
AUV SLAM and Experiments Using a Mechanical Scanning Forward-Looking Sonar
Previous Article in Special Issue
On-Chip Cellomics Assay Enabling Algebraic and Geometric Understanding of Epigenetic Information in Cellular Networks of Living Systems. 1. Temporal Aspects of Epigenetic Information in Bacteria
Open AccessArticle

Dynamics of Ras Complexes Observed in Living Cells

by Xiangyong Li 1,2, Zhiyong Cheng 3 and Honglin Jin 1,2,*
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China
MoE Key Laboratory for Biomedical Photonics, Department of Biomedical Engineering, Huazhong University of Science and Technology, Wuhan 430074, China
Wuhan Mechanical Technology College, Wuhan 430075, China
Author to whom correspondence should be addressed.
Sensors 2012, 12(7), 9411-9422;
Received: 12 June 2012 / Revised: 29 June 2012 / Accepted: 4 July 2012 / Published: 9 July 2012
(This article belongs to the Special Issue Live Cell-Based Sensors)
K-Ras works as a switch in many important intracellular signaling pathways and plays important roles in cell growth, proliferation, differentiation and carcinogenesis. For signal transduction from K-Ras to Raf1, the best-characterized effector of K-Ras, the general view is that Ras recruits Raf1 from the cytoplasm to the cell membrane. To elucidate this process, we constructed a series of fusion proteins (including Raf1 and K-Ras fused with either fluorescent proteins or fluorescent protein fragments) to compare subcellular localizations of these proteins. Bimolecular fluorescence complementation (BiFC) and a co-transfection system were used. In the BiFC system, the K-Ras/Raf1 complexes were mainly located in the cell membrane, while the Raf1 control was uniformly distributed in the cytoplasm. However, the complexes of Raf1 and K-RasC185S, a K-Ras mutant which loses membrane-localization, were also able to accumulate in the cell membrane. In contrast, an apparent cytosolic distribution pattern was observed in cells co-transfected with mcerulean-Raf1 and EGFP-K-RasC185S, suggesting that the membrane localization of K-Ras/Raf1 complexes is not entirely dependent on K-Ras, and that other factors, such as the irreversible conformation formed between K-Ras and Raf1 may play a role. This study sheds light on the interaction between K-Ras and Raf1 and provides a practical method to elucidate the mechanism underlying K-Ras and Raf1 binding to the cell membrane. View Full-Text
Keywords: K-Ras; Raf1; BiFC; membrane association; signal pathway K-Ras; Raf1; BiFC; membrane association; signal pathway
MDPI and ACS Style

Li, X.; Cheng, Z.; Jin, H. Dynamics of Ras Complexes Observed in Living Cells. Sensors 2012, 12, 9411-9422.

Show more citation formats Show less citations formats

Article Access Map by Country/Region

Only visits after 24 November 2015 are recorded.
Back to TopTop