$$^1$$ 1 H, $$^{13}$$ 13 C and $$^{15}$$ 15 N assignments of human Grb2 free of ligands
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ARTICLE
1 H, 13 C and 15 N assignments of human Grb2 free of ligands Louise Pinet1,2 · Ying‑Hui Wang1,3 · Anaïs Vogel1,4 · Françoise Guerlesquin5 · Nadine Assrir1 · Carine van Heijenoort1 Received: 7 May 2020 / Accepted: 31 July 2020 © The Author(s) 2020
Abstract Growth factor receptor-bound 2 (Grb2) is an important link in the receptor tyrosine kinase signaling cascades. It is involved in crucial processes, both physiological (mainly embryogenesis) and pathological (different types of cancer). Several binding partners of all three domains (SH3–SH2–SH3) of this adaptor protein are well described, such as ErbB family members for the SH2 domain and Sos for the SH3 domains. How the different domains interact with each other, both structurally and functionally, is still unclear. These interactions could be essential for regulation processes, and therefore are of great interest. Although a lot of structural data on Grb2 exist, they describe either individual domains, ligand-bound conformations, or frozen pictures of the protein captured by crystallography. Here we report the assignment of backbone and of 13 C𝛽 chemical shifts of full-length, apo-Grb2 in solution. In addition to the assigned conformation corresponding to three wellfolded domains, a set of peaks compatible with the presence of an unfolded conformation of the N-terminal SH3 domain is observed. This assignment paves the way for future studies of inter-domain interactions and dynamics that have to be taken into account when studying the regulation of Grb2 interactions and signaling pathways. Keywords Growth factor receptor-bound 2 (Grb2) · Adaptor protein · SH2 · SH3 · RTK signaling
Biological context Growth factor receptor-bound 2 (Grb2) is an adaptor protein essential for early steps of embryogenesis, especially differentiation (Cheng et al. 1998), but it was also shown to be upregulated in a subset of breast cancers (Daly et al. 1994). It was originally discovered in humans as the missing link between receptor tyrosine kinases (RTKs) and the Ras/ MAPK pathway (Lowenstein et al. 1992). Grb2 is 217 residues long (with a molecular weight of 25 kDa) and is made
up of three domains: a central SH2 domain, a well-described type of phosphotyrosine-binding domain, is flanked by two SH3 domains (called NSH3 at the N-terminus and CSH3 at the C-terminus) that bind to proline-rich motifs. Grb2 binds to phosphorylated RTKs via its SH2 domain and to the Son of sevenless guanine nucleotide exchange factor (Sos) via its SH3 domains, triggering Ras/MAPK pathway signaling (Lowenstein et al. 1992; Gale et al. 1993; Buday and Downward 1993), but is also involved in other signaling pathways (Gu and Neel 2003; Ravid et al. 2004; Yamazaki et al. 2002). 1
Department of Analytical and Structural Chemistry and Biology, Institut de Chimie des Substances Naturelles, CNRS UPR2301, Université Paris-Saclay, 1, av. de la terrasse, 91190 Gif‑sur‑Yvette, France
2
Present Address: Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Sw
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