title: The higher level of complexity of K-Ras4B activation at the membrane.
abstract: Is nucleotide exchange sufficient to activate K-Ras4B? To signal, oncogenic rat sarcoma (Ras) anchors in the membrane and recruits effectors by exposing its effector lobe. With the use of NMR and molecular dynamics (MD) simulations, we observed that in solution, farnesylated guanosine 5'-diphosphate (GDP)-bound K-Ras4B is predominantly autoinhibited by its hypervariable region (HVR), whereas the GTP-bound state favors an activated, HVR-released state. On the anionic membrane, the catalytic domain adopts multiple orientations, including parallel (∼180°) and perpendicular (∼90°) alignments of the allosteric helices, with respect to the membrane surface direction. In the autoinhibited state, the HVR is sandwiched between the effector lobe and the membrane; in the active state, with membrane-anchored farnesyl and unrestrained HVR, the catalytic domain fluctuates reinlessly, exposing its effector-binding site. Dimerization and clustering can reduce the fluctuations. This achieves preorganized, productive conformations. Notably, we also observe HVR-autoinhibited K-Ras4B-GTP states, with GDP-bound-like orientations of the helices. Thus, we propose that the GDP/GTP exchange may not be sufficient for activation; instead, our results suggest that the GDP/GTP exchange, HVR sequestration, farnesyl insertion, and orientation/localization of the catalytic domain at the membrane conjointly determine the active or inactive state of K-Ras4B. Importantly, K-Ras4B-GTP can exist in active and inactive states; on its own, GTP binding may not compel K-Ras4B activation.-Jang, H., Banerjee, A., Chavan, T. S, Lu, S., Zhang, J., Gaponenko, V., Nussinov, R. The higher level of complexity of K-Ras4B activation at the membrane.
authors.affiliation: *Basic Science Program, Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Cancer and Inflammation Program, National Cancer Institute at Frederick, Frederick, Maryland, USA; Department of Chemistry, Department of Medicinal Chemistry, and Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois, USA; Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis, Chinese Ministry of Education, Shanghai JiaoTong University, School of Medicine, Shanghai, China; and Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Hyunbum
authors.initials: H
authors.lastname: Jang
authors.affiliation: *Basic Science Program, Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Cancer and Inflammation Program, National Cancer Institute at Frederick, Frederick, Maryland, USA; Department of Chemistry, Department of Medicinal Chemistry, and Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois, USA; Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis, Chinese Ministry of Education, Shanghai JiaoTong University, School of Medicine, Shanghai, China; and Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Avik
authors.initials: A
authors.lastname: Banerjee
authors.affiliation: *Basic Science Program, Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Cancer and Inflammation Program, National Cancer Institute at Frederick, Frederick, Maryland, USA; Department of Chemistry, Department of Medicinal Chemistry, and Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois, USA; Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis, Chinese Ministry of Education, Shanghai JiaoTong University, School of Medicine, Shanghai, China; and Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Tanmay S
authors.initials: TS
authors.lastname: Chavan
authors.affiliation: *Basic Science Program, Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Cancer and Inflammation Program, National Cancer Institute at Frederick, Frederick, Maryland, USA; Department of Chemistry, Department of Medicinal Chemistry, and Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois, USA; Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis, Chinese Ministry of Education, Shanghai JiaoTong University, School of Medicine, Shanghai, China; and Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Shaoyong
authors.initials: S
authors.lastname: Lu
authors.affiliation: *Basic Science Program, Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Cancer and Inflammation Program, National Cancer Institute at Frederick, Frederick, Maryland, USA; Department of Chemistry, Department of Medicinal Chemistry, and Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois, USA; Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis, Chinese Ministry of Education, Shanghai JiaoTong University, School of Medicine, Shanghai, China; and Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.affiliation: *Basic Science Program, Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Cancer and Inflammation Program, National Cancer Institute at Frederick, Frederick, Maryland, USA; Department of Chemistry, Department of Medicinal Chemistry, and Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois, USA; Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis, Chinese Ministry of Education, Shanghai JiaoTong University, School of Medicine, Shanghai, China; and Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel nussinor@helix.nih.gov vadimg@uic.edu.
authors.forename: Vadim
authors.initials: V
authors.lastname: Gaponenko
authors.affiliation: *Basic Science Program, Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Cancer and Inflammation Program, National Cancer Institute at Frederick, Frederick, Maryland, USA; Department of Chemistry, Department of Medicinal Chemistry, and Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois, USA; Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis, Chinese Ministry of Education, Shanghai JiaoTong University, School of Medicine, Shanghai, China; and Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel nussinor@helix.nih.gov vadimg@uic.edu.
authors.forename: Ruth
authors.initials: R
authors.lastname: Nussinov
mesh_terms: D000818:Animals; D020134:Catalytic Domain; D002462:Cell Membrane / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000737:chemistry / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D056004:Molecular Dynamics Simulation; D015394:Molecular Structure; D041681:NIH 3T3 Cells; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D016283:Proto-Oncogene Proteins p21(ras) / Q000737:chemistry / Q000378:metabolism*
chemical_list: D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; C495971:Hras protein, mouse; D016283:Proto-Oncogene Proteins p21(ras)
keywords: GDP/GTP exchange; KRAS; farnesyl insertion; phospholipids; signaling