TitleMultivalent Angiomotin-like 1 and Yes-associated protein form a dynamic complex.
Publication TypeJournal Article
Year of Publication2022
AuthorsVogel A, Crawford A, Nyarko A
JournalProtein Sci
Volume31
Issue5
Paginatione4295
Date Published2022 05
ISSN1469-896X
KeywordsAngiomotins, Binding Sites, Peptides, Transcription Factors, YAP-Signaling Proteins
Abstract

Multivalent complexes formed between the cancer-promoting transcriptional co-activator, Yes-associated protein (YAP), and proteins containing short linear motifs of type PPxY modulate cell proliferation and are attractive therapeutic targets. However, challenges producing PPxY polypeptides containing the full binding domain has limited understanding of the assembly process. Here, we successfully produced a polypeptide containing the complete set of three PPxY binding sites of Angiomotin-like 1 (AMOTL1), a scaffolding protein that regulates the nucleo-cytoplasmic shuttling of YAP via WW-PPxY interactions. Using an array of biophysical techniques including isothermal titration calorimetry, size-exclusion chromatography coupled to multi-angle light scattering, and solution nuclear magnetic resonance spectroscopy, we show that the AMOTL1 polypeptide is partially disordered, and binds the YAP WW domains to form an ensemble of complexes of varying stabilities. The binding process is initiated by the binding of one YAP WW domain to one AMOTL1 PPxY motif and is completed by transient interactions of the second YAP WW domain with a second AMOTL1 PPxY motif to form an equilibrating mixture composed of various species having two YAP sites bound to two conjugate AMOTL1 sites. We rationalize that the transient interactions fine-tune the stability of the complex for rapid assembly and disassembly in response to changes in the local cellular environment.

DOI10.1002/pro.4295
Alternate JournalProtein Sci
PubMed ID35481651
PubMed Central IDPMC8994507
Grant List1S10OD018518 / NH / NIH HHS / United States