Biochemical and structural analysis reveals the Neurofibromin (NF1) protein forms a high-affinity dimer

Sherekar, M.; Han, S.-W.; Ghirlando, R.; Messing, S.; Drew, M.; Juneja, P.; O'Neill, H.; Stanley, C.; Bhowmik, D.; Ramanathan, A.; Subramaniam, S.; Nissley, D. V.; Gillette, W.; McCormick, F.; Esposito, D.

doi:10.1101/757856

bioRxiv · 10.1101/757856

Biochemical and structural analysis reveals the Neurofibromin (NF1) protein forms a high-affinity dimer

Sherekar, M.·Han, S.-W.·Ghirlando, R.·Messing, S.·Drew, M.·Juneja, P.·O'Neill, H.·Stanley, C.·Bhowmik, D.·Ramanathan, A.·Subramaniam, S.·Nissley, D. V.·Gillette, W.·McCormick, F.·Esposito, D.

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Abstract

Neurofibromin is the protein product of the NF1 gene which is mutated in the Rasopathy disease Neurofibromatosis Type I. Defects in NF1 lead to aberrant signaling through the RAS-MAPK pathway due to disruption of the Neurofibromin GTPase-activating function on RAS family small GTPases. Very little is known about the function of the majority of Neurofibromin--to date, biochemical and structural data exist only for the GAP domain and the region containing a Sec-PH motif. To better understand the role of this large protein, we carried out a series of biochemical and biophysical studies which demonstrate that full length Neurofibromin forms a high-affinity dimer. Neurofibromin dimerization also occurs in cells, and likely has biological and clinical implications. Analysis of purified full-length and truncated variants of Neurofibromin by negative stain electron microscopy reveals the overall architecture of the dimer and predicts the potential interactions which contribute to the dimer interface. Structures resembling high-affinity full-length dimers could be reconstituted by mixing N- and C-terminal protein domains in vitro. Taken together these data suggest how Neurofibromin dimers might form and be stabilized within the cell.

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Sherekar, M., Han, S.-W., Ghirlando, R., Messing, S., Drew, M., Juneja, P., O'Neill, H., Stanley, C., Bhowmik, D., Ramanathan, A., Subramaniam, S., Nissley, D. V., Gillette, W., McCormick, F., Esposito, D.. 2019-09-05. Biochemical and structural analysis reveals the Neurofibromin (NF1) protein forms a high-affinity dimer. https://doi.org/10.1101/757856

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