Crystal structure of the RUN domain of the RAP2-interacting protein x
JOURNAL OF BIOLOGICAL CHEMISTRY
Authors: Kukimoto-Niino, Mutsuko; Takagi, Tetsuo; Akasaka, Ryogo; Murayama, Kazutaka; Uchikubo-Kamo, Tomomi; Terada, Takaho; Inoue, Makoto; Watanabe, Satoru; Tanaka, Akiko; Hayashizaki, Yoshihide; Kigawa, Takanori; Shirouzu, Mikako; Yokoyama, Shigeyuki
Abstract
Rap2-interacting protein x (RPIPx) is a homolog of RPIP8, a specific effector of Rap2 GTPase. The N-terminal region of RPIP8, which contains the RUN domain, interacts with Rap2. Using cell-free synthesis and NMR, we determined that the region encompassing residues 83-255 of mouse RPIPx, which is 40-residues larger than the predicted RUN domain (residues 113-245), is the minimum fragment that forms a correctly folded protein. This fragment, the RPIPx RUN domain, interacted specifically with Rap2B in vitro in a nucleotide-dependent manner. The crystal structure of the RPIPx RUN domain was determined at 2.0 angstrom of resolution by the multiwavelength anomalous dispersion (MAD) method. The RPIPx RUN domain comprises eight anti-parallel alpha-helices, which form an extensive hydrophobic core, followed by an extended segment. The residues in the core region are highly conserved, suggesting the conservation of the RUN domain-fold among the RUN domain-containing proteins. The residues forming a positively charged surface are conserved between RPIP8 and its homologs, suggesting that this surface is important for Rap2 binding. In the crystal the putative Rap2 binding site of the RPIPx RUN domain interacts with the extended segment in a segment-swapping manner.
Rap2b promotes proliferation, migration, and invasion of lung cancer cells
JOURNAL OF RECEPTORS AND SIGNAL TRANSDUCTION
Authors: Peng, Yi-Gen; Zhang, Zheng-Qun; Chen, Yan-bin; Huang, Jian-An
Abstract
Rap2b, a member of the guanosine triphosphate-binding proteins, is widely up-regulated in many types of tumors. However, the functional role of Rap2b in tumorigenesis of lung cancer remains to be fully elucidated. In this study, we investigated the effect of Rap2b on the lung cancer malignant phenotype, such as cell proliferation and metastasis. We found that Rap2b could promote the abilities of lung cancer cell wound healing, migration, and invasion via increasing matrix metalloproteinase-2 enzyme activity. Furthermore, Rap2b overexpression could increase the phosphorylation level of extracellular signal-regulated protein kinases 1/2. In conclusion, our results suggested that Rap2b may be a potential therapeutic target for lung cancer.