1 H, 13 C, and 15 N chemical shift assignment of human PACSIN1/syndapin I SH3 domain in solution
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ARTICLE
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H, 13C, and 15N chemical shift assignment of human PACSIN1/ syndapin I SH3 domain in solution Emmanuelle Boll1,2 · Francois‑Xavier Cantrelle1,2 · Isabelle Landrieu1,2 · Matthieu Hirel1,2 · Davy Sinnaeve1,2 · Géraldine Levy1,2 Received: 20 January 2020 / Accepted: 26 March 2020 © Springer Nature B.V. 2020
Abstract Human neuron-specific PACSIN1 plays a key role in synaptic vesicle recycling and endocytosis, as well as reorganization of the microtubule dynamics to maintain axonal plasticity. PACSIN1 contains a highly conserved C-terminal SH3 domain and an F-bar domain at its N-terminus. Due to its remarkable interaction network, PACSIN1 plays a central role in key neuronal functions. Here, we present a robust backbone and side-chain assignment of PACSIN1 SH3 domain based on 2D [ 1H,15N] HSQC or HMQC, and 3D BEST-HNCO, -HNCACB, -HN(CO)CACB, -HN(CA)CO, and standard (H)CC(CO)NH, HN(CA) NNH, HN(COCA)NH, HBHANNH, HNHA, HBHA(CO)NH, H(CC)(CO)NH, HCCH-TOCSY, that covers 96% for all 13CO, 13 Cα and 13Cβ, 28% of 13Cγδε, and 95% of 1HN and 15N chemical shifts. Modelling based on sequence homology with a known related structure, and chemical shift-based secondary structure predictions, identified the presence of five β-strands linked by flexible loops. Taken together, these results open up new avenues to investigate and develop new therapeutic strategies. Keywords PACSIN1 · SH3 domain · NMR resonance assignment · Protein–protein interaction
Biological context The Human neurospecific PACSIN1 or Phosphoprotein Protein Kinase C and Casein Kinase substrate in neurons protein 1, which is also known as syndapin I (synaptic dynamin-associated protein), plays a central role in synaptic vesicle recycling and endocytosis, and reorganization of the microtubule and actin cytoskeleton (Modregger et al. 2002; Qualmann et al. 1999; Anggono et al. 2006). PACSIN1 comprised of 458 amino acid residues is typically found along neurites and within the synaptic boutons. In eukaryotes, PACSIN1 has two isoforms, PACSIN2 and 3, showing a wide tissue distribution (Modregger et al. 2000). PACSIN2 Emmanuelle Boll and Francois-Xavier Cantrelle have contributed equally to this work. * Géraldine Levy geraldine.levy@univ‑lille.fr 1
Institut Pasteur de Lille, U1167 ‑ RID‑AGE ‑ Risk Factors and Molecular Determinants of Aging‑Related Diseases, Univ. Lille, Inserm, CHU Lille, 59000 Lille, France
CNRS ERL Integrative Structural Biology, 59000 Lille, France
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is ubiquitously expressed, PACSIN3 is present mainly in skeletal muscles, heart and lung, while PACSIN1 is found exclusively in the Central Nervous System (CNS). The divergence of the three isoforms spatial expression mirrors fine-tuning in their specificity in recruitment of other proteins involved in endocytosis (Modregger et al. 2000; Kessels and Qualmann 2004). All isoforms share a highly conserved Fer-CIP4 homology-BAR (F-BAR) at its N-terminus (Qualmann et al. 2011), which consists of a 3-helix bundle and has been shown to mediate and/or stabilize membrane curvature (Rao et al. 2
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