{"doi":"10.1101/2025.09.11.675621","title":"Effects of Nanopore Confinement on the Conformational, Dynamical, and Self-Assembly Properties of an FG-Repeat Peptide","abstract":"ABSTRACT The central channels of nuclear pore complexes (NPCs) in eukaryotic cells are filled with protein chains whose sequences contain characteristic phenylalanine-glycine motifs. Properties of these nanopore-confined FG-repeat sequences are of central importance to NPC function. Here we demonstrate an approach to nuclear magnetic resonance (NMR) studies of FG-repeat sequences (or other polypeptides) that are tethered within pores with diameters similar to those of NPC channels. By attaching alkyl phosphonate groups to the N-terminus of a 30-residue peptide that contains four FG repeats, called FG30, we tether FG30 chains to walls of 20-nm-diameter pores in anodic aluminum oxide (AAO) wafers through phosphonate-surface bonds. Quantitative 13 C and 31 P NMR measurements indicate 90 mM peptide concentrations (300 mg/ml) within the pores. NMR spectra and spin relaxation measurements show that FG30 chains are dynamically disordered and random-coil-like in buffer-filled pores over a broad temperature range. In contrast, FG30 aggregates in free solution at concentrations above 2 mM, forming structurally ordered fibrils according to electron microscopy and NMR measurements. These results demonstrate the utility of AAO as a scaffold for studies of polypeptides in nanopore-confined environments and show that tethering to nanopore walls can dramatically alter the self-assembly properties of an FG-repeat sequence. TOC Graphic","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":559079,"datarank":0.10397207708399181,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"self_citation_contribution":0.10397207708399181,"citation_network_contribution":0.0,"self_endowment_contribution":0.10397207708399181,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9499,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":337240,"name":"Wai‐Ming Yau","orcid":"0000-0002-7668-7780","position":1,"is_corresponding":false},{"id":272110,"name":"Robert Tycko","orcid":"0000-0001-7039-7275","position":2,"is_corresponding":false},{"id":649469,"name":"Wancheng Zhao","orcid":"0000-0001-6727-5533","position":0,"is_corresponding":true}],"reference_count":76,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:55:30.312295Z","pmid":"41019633","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}