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Second Harmonic Generation Detection of Ras Conformational Changes and Discovery of a Small Molecule Binder

Second Harmonic Generation Detection of Ras Conformational Changes and Discovery of a Small Molecule Binder

Elizabeth Donohue, Sina Khorsand, Gabriel Mercado, Kristen M Varney, Paul T Wilder, Wenbo Yu, Alexander D MacKerell Jr, Patrick Alexander, Que N Van, Ben Moree, Andrew G Stephen, David J Weber, Joshua Salafsky, etc.

Proc Natl Acad Sci U S A. 2019 Aug 27;116(35):17290-17297.

PMID: 31399543

Abstract:

Second harmonic generation (SHG) is an emergent biophysical method that sensitively measures real-time conformational change of biomolecules in the presence of biological ligands and small molecules. This study describes the successful implementation of SHG as a primary screening platform to identify fragment ligands to oncogenic Kirsten rat sarcoma (KRas). KRas is the most frequently mutated driver of pancreatic, colon, and lung cancers; however, there are few well-characterized small molecule ligands due to a lack of deep binding pockets. Using SHG, we identified a fragment binder to KRasG12D and used 1H 15N transverse relaxation optimized spectroscopy (TROSY) heteronuclear single-quantum coherence (HSQC) NMR to characterize its binding site as a pocket adjacent to the switch 2 region. The unique sensitivity of SHG furthered our study by revealing distinct conformations induced by our hit fragment compared with 4,6-dichloro-2-methyl-3-aminoethyl-indole (DCAI), a Ras ligand previously described to bind the same pocket. This study highlights SHG as a high-throughput screening platform that reveals structural insights in addition to ligand binding.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP299165927	DCAI		299165-92-7	Price

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