Language

English

Publication Date

5-7-2024

Journal

Biochemistry

DOI

10.1021/acs.biochem.4c00056

PMID

38619104

PMCID

PMC12022813

PubMedCentral® Posted Date

4-25-2025

PubMedCentral® Full Text Version

Author MSS

Abstract

NANOG protein levels correlate with stem cell pluripotency. NANOG concentrations fluctuate constantly with low NANOG levels leading to spontaneous cell differentiation. Previous literature implicated Pin1, a phosphorylation-dependent prolyl isomerase, as a key player in NANOG stabilization. Here, using NMR spectroscopy, we investigate the molecular interactions of Pin1 with the NANOG unstructured N-terminal domain that contains a PEST sequence with two phosphorylation sites. Phosphorylation of NANOG PEST peptides increases affinity to Pin1. By systematically increasing the amount of cis PEST conformers, we show that the peptides bind tighter to the prolyl isomerase domain (PPIase) of Pin1. Phosphorylation and cis Pro enhancement at both PEST sites lead to a 5–10-fold increase in NANOG binding to the Pin1 WW domain and PPIase domain, respectively. The cis-populated NANOG PEST peptides can be potential inhibitors for disrupting Pin1-dependent NANOG stabilization in cancer stem cells.

Keywords

NIMA-Interacting Peptidylprolyl Isomerase, Nanog Homeobox Protein, Phosphorylation, Humans, Protein Stability, Protein Binding, Stereoisomerism

Published Open-Access

yes

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