GORK K+ channel structure and gating vital to informing stomatal engineering

• Published in: Nature communications Vol. 16; no. 1; pp. 1961 - 13
• Main Authors: Zhang, Xue, Carroll, William, Nguyen, Thu Binh-Anh, Nguyen, Thanh-Hao, Yang, Zhao, Ma, Miaolian, Huang, Xiaowei, Hills, Adrian, Guo, Hui, Karnik, Rucha, Blatt, Michael R., Zhang, Peng
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 25.02.2025; Nature Publishing Group; Nature Portfolio
• Subjects: 101/28; 631/1647/1453/1971; 631/1647/2258/1258/1259; 631/449/2124; 82/80; 82/83; 9/74; Angiosperms; Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis Proteins - chemistry; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Bioengineering; Carbon sequestration; Channel gating; Cyclic nucleotides; Efflux; Electric potential; Functional analysis; Homology; Humanities and Social Sciences; Ion Channel Gating - physiology; Models, Molecular; multidisciplinary; Nucleotides; Physiology; Plant Stomata - metabolism; Plant Stomata - physiology; Potassium; Potassium - metabolism; Potassium channels; Potassium Channels - chemistry; Potassium Channels - genetics; Potassium Channels - metabolism; Protein Conformation; Protein Domains; Proteins; Science; Science (multidisciplinary); Sensors; Stomata; Structure-function relationships; Symmetry; Voltage; Water use; Water use efficiency
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-025-57287-7

The Arabidopsis GORK channel is a major pathway for guard cell K + efflux that facilitates stomatal closure. GORK is an outwardly-rectifying member of the cyclic-nucleotide binding-homology domain (CNBHD) family of K + channels with close homologues in all other angiosperms known to date. Its bioengineering has demonstrated the potential for enhanced carbon assimilation and water use efficiency. Here we identify critical domains through structural and functional analysis, highlighting conformations that reflect long-lived closed and pre-open states of GORK. These conformations are marked by interactions at the cytosolic face of the membrane between so-called voltage-sensor, C-linker and CNBHD domains, the latter relocating across 10 Å below the voltage sensor. The interactions center around two coupling sites that functional analysis establish are critical for channel gating. The channel is also subject to putative, ligand-like interactions within the CNBHD, which leads to its gating independence of cyclic nucleotides such as cAMP or cGMP. These findings implicate a multi-step mechanism of semi-independent conformational transitions that underlie channel activity and offer promising new sites for optimizing GORK to engineer stomata. This study identifies long-lived closed and pre-open states of GORK K+ channel through structural and functional analysis, suggesting semi-independent conformational transitions underlying channel activity and promising sites for stomata engineering.