Barley Nodulin 26-like intrinsic protein permeates water, metalloids, saccharides, and ion pairs due to structural plasticity and diversification

• Published in: The Journal of biological chemistry Vol. 299; no. 12; p. 105410
• Main Authors: Venkataraghavan, Akshayaa, Schwerdt, Julian G., Tyerman, Stephen D., Hrmova, Maria
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.2023; American Society for Biochemistry and Molecular Biology
• Subjects: 3D molecular modeling of aquaporins; Animals; Aquaporins - metabolism; Glucosamine; heterologous protein expression; Hordeum - metabolism; Metalloids - metabolism; phylogenomics; solute permeation selectivity; stopped-flow spectrophotometry; Sucrose; Water - metabolism; stopped-flow spectrophotometry; UniProt; Co; FAAR; EBI; 3D molecular modeling of aquaporins; PDB; P; MEME; HvNIP2;1; NOD26; NCBI; AQP(s); DMPC; heterologous protein expression; phylogenomics; FHTR; Vw; aqpN; IMAC; solute permeation selectivity; SMA; HIP; GIP; HEK; SoPIP2;1; SDS-PAGE; NIP; GSGR; HEPES; PIP; TIP; SIP; NPA; XIP; BA
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/j.jbc.2023.105410

Aquaporins can facilitate the passive movement of water, small polar molecules, and some ions. Here, we examined solute selectivity for the barley Nodulin 26-like Intrinsic Protein (HvNIP2;1) embedded in liposomes and examined through stopped-flow light scattering spectrophotometry and Xenopus laevis oocyte swelling assays. We found that HvNIP2;1 permeates water, boric and germanic acids, sucrose, and lactose but not d-glucose or d-fructose. Other saccharides, such as neutral (d-mannose, d-galactose, d-xylose, d-mannoheptaose) and charged (N-acetyl d-glucosamine, d-glucosamine, d-glucuronic acid) aldoses, disaccharides (cellobiose, gentiobiose, trehalose), trisaccharide raffinose, and urea, glycerol, and acyclic polyols, were permeated to a much lower extent. We observed apparent permeation of hydrated KCl and MgSO4 ions, while CH3COONa and NaNO3 permeated at significantly lower rates. Our experiments with boric acid and sucrose revealed no apparent interaction between solutes when permeated together, and AgNO3 or H[AuCl4] blocked the permeation of all solutes. Docking of sucrose in HvNIP2;1 and spinach water-selective SoPIP2;1 aquaporins revealed the structural basis for sucrose permeation in HvNIP2;1 but not in SoPIP2;1, and defined key residues interacting with this permeant. In a biological context, sucrose transport could constitute a novel element of plant saccharide-transporting machinery. Phylogenomic analyses of 164 Viridiplantae and 2993 Archaean, bacterial, fungal, and Metazoan aquaporins rationalized solute poly-selectivity in NIP3 sub-clade entries and suggested that they diversified from other sub-clades to acquire a unique specificity of saccharide transporters. Solute specificity definition in NIP aquaporins could inspire developing plants for food production.