Disorder in a two-domain neuronal Ca2+-binding protein regulates domain stability and dynamics using ligand mimicry

Lasse Staby, Katherine R. Kemplen, Amelie Stein, Michael Ploug, Jane Clarke, Karen Skriver, Pétur O. Heidarsson, Birthe B. Kragelund*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Understanding the interplay between sequence, structure and function of proteins has been complicated in recent years by the discovery of intrinsically disordered proteins (IDPs), which perform biological functions in the absence of a well-defined three-dimensional fold. Disordered protein sequences account for roughly 30% of the human proteome and in many proteins, disordered and ordered domains coexist. However, few studies have assessed how either feature affects the properties of the other. In this study, we examine the role of a disordered tail in the overall properties of the two-domain, calcium-sensing protein neuronal calcium sensor 1 (NCS-1). We show that loss of just six of the 190 residues at the flexible C-terminus is sufficient to severely affect stability, dynamics, and folding behavior of both ordered domains. We identify specific hydrophobic contacts mediated by the disordered tail that may be responsible for stabilizing the distal N-terminal domain. Moreover, sequence analyses indicate the presence of an LSL-motif in the tail that acts as a mimic of native ligands critical to the observed order–disorder communication. Removing the disordered tail leads to a shorter life-time of the ligand-bound complex likely originating from the observed destabilization. This close relationship between order and disorder may have important implications for how investigations into mixed systems are designed and opens up a novel avenue of drug targeting exploiting this type of behavior.

Original languageEnglish
Pages (from-to)2263-2278
Number of pages16
JournalCellular and Molecular Life Sciences
Issue number5
Publication statusPublished - 16 Sept 2020

Bibliographical note

Funding Information:
We thank Signe Sjørup for DNA constructs, Jacob Hertz Martinsen for help with protein purification, and Gry Ellis Rasmussen for running the SPR experiments. This work was supported by the Lundbeck Foundation (to K.K., P.O.H., and A.S.), the Novo Nordisk Foundation (P.O.H.), the Novo Nordisk Foundation Challenge program (grant #NNF18OC0033926)—REPIN, rethinking protein interactions (to B.B.K. and K.S.), and the Danish Council for Independent Research, Natural Sciences (4181-00344 to B.B.K. and K.S). We thank VILLUMFONDEN for generous support for NMR equipment.

Publisher Copyright:
© 2020, The Author(s).

Other keywords

  • EF-hand
  • Frequenin
  • IDP
  • NCS-1
  • Order–disorder interplay
  • Protein folding


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