Fig. 1: Mb24 specifically interacts with NRAS.
From: Inhibition and degradation of NRAS with a pan-NRAS monobody
A Biolayer interferometry (BLI) sensorgrams of RAS isotypes loaded with either GTPγS or GDP to Mb24 immobilized on a sensor tip. The KD values shown are from global fitting of a 1:1 binding model. N.D. not determined due to weak binding. B Mb24 and NS1 share an overlapping epitope. BLI sensorgram of immobilized Mb24 binding to NRAS(K135R)•GDP binding followed by the addition of NS1 (left graph). NS1 did not bind to the NRAS(K135R) precomplexed with Mb24. NS1 does bind to immobilized NRAS(K135R) in the absence of Mb24 (right graph). C Colocalization of mCherry-tagged Mb24 with EGFP-tagged KRAS4B or NRAS in cotransfected HEK 293 cells. The graphs show the fluorescence intensity profiles across the microscopy images. D Coimmunoprecipitation of Mbs from HEK 293 cells cotransfected with FLAG-tagged CFP alone, CPF-NS1, or CFP-Mb24 and HA-tagged HRASG12V, KRASG12V, or NRASG12V. Top panel, αFLAG immunoprecipitates were probed with αFLAG and αHA antibodies. Bottom panels, whole cell lysates (WCL) were probed with the indicated antibodies.
