The complex structure of ARAP3 RhoGAP-RhoA reveals the molecular mechanism for specifically recognition of RhoA by ARAP3
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摘要
ARAP3 is unique for its dual specific GAPs(GTPase activating protein) activity for Arf6 and RhoA, regulated by Ptd Ins(3,4,5)P3 and Rap1-GTP and involving in regulation of cell shape and adhesion. However, the clear interface between the Rho GAP domain and RhoA is unknown. Meanwhile, the substrate selectivity of the Rho GAP domain is also unclear. In this study, we solved the crystal structure of RhoA in complex with the Rho GAP domain of ARAP3. The complex structure presented the clear interface between the Rho GAP domain and RhoA. By analyzing the complex structure in combination with in vitro RhoA activity assay and ITC experiments, we found the crucial residues affecting the Rho GAP activity and the substrates selectivity among RhoA, Rac1 and Cdc42. The title should be in bold Arial(14 points), with the initial letter capitalized. Authors will start on a new line and should be in regular Times New Roman(10 points). Family name should be placed after the first name. Underline the name of the presenting author. Affiliation of authors should start on a new line in italics with address and e-mail address. Main text should be within 400 words in regular Times New Roman(12 points) and single-spaced. Insert a single blank line space between paragraphs.
ARAP3 is unique for its dual specific GAPs(GTPase activating protein) activity for Arf6 and RhoA, regulated by Ptd Ins(3,4,5)P3 and Rap1-GTP and involving in regulation of cell shape and adhesion. However, the clear interface between the Rho GAP domain and RhoA is unknown. Meanwhile, the substrate selectivity of the Rho GAP domain is also unclear. In this study, we solved the crystal structure of RhoA in complex with the Rho GAP domain of ARAP3. The complex structure presented the clear interface between the Rho GAP domain and RhoA. By analyzing the complex structure in combination with in vitro RhoA activity assay and ITC experiments, we found the crucial residues affecting the Rho GAP activity and the substrates selectivity among RhoA, Rac1 and Cdc42. The title should be in bold Arial(14 points), with the initial letter capitalized. Authors will start on a new line and should be in regular Times New Roman(10 points). Family name should be placed after the first name. Underline the name of the presenting author. Affiliation of authors should start on a new line in italics with address and e-mail address. Main text should be within 400 words in regular Times New Roman(12 points) and single-spaced. Insert a single blank line space between paragraphs.
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