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Pathogenic Mutations Associated with Legius Syndrome Modify the Spred1 Surface and Are Involved in Direct Binding to the Ras Inactivator Neurofibromin.
J Mol Biol. 2019 09 06; 431(19):3889-3899.JM

Abstract

Neurofibromatosis type I (NF1) and Legius syndrome are rare inherited disorders that share diagnostic symptoms including dermal abnormalities like axillary and inguinal freckling and café au lait spots. In addition, patients suffering from NF1 have a demanding risk for the development of severe tumors of the peripheral and central nervous system among other NF1-specific symptoms. NF1 and Legius syndrome are caused by alterations in the NF1 and SPRED1 genes encoding the Ras inhibitors neurofibromin and Spred1 (sprouty related EVH1 domain-containing protein), respectively. Neurofibromin functions as a Ras-specific GTPase-activating protein (Ras-GAP), and Spred1 enhances Ras inactivation by recruiting neurofibromin from the cytosol to membrane-anchored Ras. In a previous study, we mapped the Spred binding site to the GAP-related domain of neurofibromin (NF1-GAP) and identified the GAPex subdomain as critical for Spred1 binding. Here, we characterize the binding site of these proteins in more detail focusing on a mutant Spred1 variant carrying a pathogenic missense mutation (threonine 102 to arginine). Introduction of this mutation, which locates at the N-terminal EVH1 domain of Spred1, weakens the interaction with neurofibromin by about 3 orders of magnitude without perturbing the protein fold, and the binding site of NF1-GAP on the mutant Spred1(EVH1) variant can be identified by NMR spectroscopy. Taken together, our data provide structural insight into the interaction of Spred1 and neurofibromin and characterize the structural or functional consequence of selected patient-derived mutations associated with Legius syndrome.

Authors+Show Affiliations

Division of Biological Chemistry, Biocenter, Medical University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria; Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.Division of Biological Chemistry, Biocenter, Medical University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria. Electronic address: theresia.dunzendorfer-matt@i-med.ac.at.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

31401120

Citation

Führer, Sebastian, et al. "Pathogenic Mutations Associated With Legius Syndrome Modify the Spred1 Surface and Are Involved in Direct Binding to the Ras Inactivator Neurofibromin." Journal of Molecular Biology, vol. 431, no. 19, 2019, pp. 3889-3899.
Führer S, Tollinger M, Dunzendorfer-Matt T. Pathogenic Mutations Associated with Legius Syndrome Modify the Spred1 Surface and Are Involved in Direct Binding to the Ras Inactivator Neurofibromin. J Mol Biol. 2019;431(19):3889-3899.
Führer, S., Tollinger, M., & Dunzendorfer-Matt, T. (2019). Pathogenic Mutations Associated with Legius Syndrome Modify the Spred1 Surface and Are Involved in Direct Binding to the Ras Inactivator Neurofibromin. Journal of Molecular Biology, 431(19), 3889-3899. https://doi.org/10.1016/j.jmb.2019.07.038
Führer S, Tollinger M, Dunzendorfer-Matt T. Pathogenic Mutations Associated With Legius Syndrome Modify the Spred1 Surface and Are Involved in Direct Binding to the Ras Inactivator Neurofibromin. J Mol Biol. 2019 09 6;431(19):3889-3899. PubMed PMID: 31401120.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Pathogenic Mutations Associated with Legius Syndrome Modify the Spred1 Surface and Are Involved in Direct Binding to the Ras Inactivator Neurofibromin. AU - Führer,Sebastian, AU - Tollinger,Martin, AU - Dunzendorfer-Matt,Theresia, Y1 - 2019/08/08/ PY - 2019/06/03/received PY - 2019/07/29/revised PY - 2019/07/31/accepted PY - 2019/8/12/pubmed PY - 2020/6/17/medline PY - 2019/8/12/entrez KW - NMR resonance assignment KW - Ras/MAPK pathway KW - protein interaction KW - recombinant protein KW - surface plasmon resonance SP - 3889 EP - 3899 JF - Journal of molecular biology JO - J Mol Biol VL - 431 IS - 19 N2 - Neurofibromatosis type I (NF1) and Legius syndrome are rare inherited disorders that share diagnostic symptoms including dermal abnormalities like axillary and inguinal freckling and café au lait spots. In addition, patients suffering from NF1 have a demanding risk for the development of severe tumors of the peripheral and central nervous system among other NF1-specific symptoms. NF1 and Legius syndrome are caused by alterations in the NF1 and SPRED1 genes encoding the Ras inhibitors neurofibromin and Spred1 (sprouty related EVH1 domain-containing protein), respectively. Neurofibromin functions as a Ras-specific GTPase-activating protein (Ras-GAP), and Spred1 enhances Ras inactivation by recruiting neurofibromin from the cytosol to membrane-anchored Ras. In a previous study, we mapped the Spred binding site to the GAP-related domain of neurofibromin (NF1-GAP) and identified the GAPex subdomain as critical for Spred1 binding. Here, we characterize the binding site of these proteins in more detail focusing on a mutant Spred1 variant carrying a pathogenic missense mutation (threonine 102 to arginine). Introduction of this mutation, which locates at the N-terminal EVH1 domain of Spred1, weakens the interaction with neurofibromin by about 3 orders of magnitude without perturbing the protein fold, and the binding site of NF1-GAP on the mutant Spred1(EVH1) variant can be identified by NMR spectroscopy. Taken together, our data provide structural insight into the interaction of Spred1 and neurofibromin and characterize the structural or functional consequence of selected patient-derived mutations associated with Legius syndrome. SN - 1089-8638 UR - https://www.unboundmedicine.com/medline/citation/31401120/Pathogenic_Mutations_Associated_with_Legius_Syndrome_Modify_the_Spred1_Surface_and_Are_Involved_in_Direct_Binding_to_the_Ras_Inactivator_Neurofibromin_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(19)30500-5 DB - PRIME DP - Unbound Medicine ER -