Miller_Turner_NatMed2019_nihms-1521224.pdf (1.95 MB)
Download file

Intracellular MLCK1 diversion reverses barrier loss to restore mucosal homeostasis

Download (1.95 MB)
journal contribution
posted on 07.01.2022, 20:20 by WV Graham, W He, AM Marchiando, J Zha, G Singh, HS Li, A Biswas, MLDM Ong, ZH Jiang, W Choi, H Zuccola, Y Wang, J Griffith, J Wu, HJ Rosenberg, SB Snapper, D Ostrov, SC Meredith, Lawrence MillerLawrence Miller, JR Turner
Epithelial barrier loss is a driver of intestinal and systemic diseases. Myosin light chain kinase (MLCK) is a key effector of barrier dysfunction and a potential therapeutic target, but enzymatic inhibition has unacceptable toxicity. Here, we show that a unique domain within the MLCK splice variant MLCK1 directs perijunctional actomyosin ring (PAMR) recruitment. Using the domain structure and multiple screens, we identify a domain-binding small molecule (divertin) that blocks MLCK1 recruitment without inhibiting enzymatic function. Divertin blocks acute, tumor necrosis factor (TNF)-induced MLCK1 recruitment as well as downstream myosin light chain (MLC) phosphorylation, barrier loss, and diarrhea in vitro and in vivo. Divertin corrects barrier dysfunction and prevents disease development and progression in experimental inflammatory bowel disease. Beyond applications of divertin in gastrointestinal disease, this general approach to enzymatic inhibition by preventing access to specific subcellular sites provides a new paradigm for safely and precisely targeting individual properties of enzymes with multiple functions.

Funding

Targeted Lanthanide Contrast Agents for in Cellulo Single Molecule Imaging | Funder: National Institutes of Health (National Institute of General Medical Sciences) | Grant ID: R01GM081030

HTS Assays for Inhibitors of Protien Interactions Including MLCK1-FKBP8. | Funder: National Institutes of Health (National Institute of Diabetes and Digestive and Kidney Diseases) | Grant ID: R56DK094954

Multiplexed FRET Imaging of Protein Interactions with Lanthanide Probes (EQUIPMENT SUPPLEMENT) | Funder: National Institutes of Health (National Institute of General Medical Sciences) | Grant ID: R01GM081030

History

Citation

Graham, W. V., He, W., Marchiando, A. M., Zha, J., Singh, G., Li, H. S., Biswas, A., Ong, M. L. D. M., Jiang, Z. H., Choi, W., Zuccola, H., Wang, Y., Griffith, J., Wu, J., Rosenberg, H. J., Wang, Y., Snapper, S. B., Ostrov, D., Meredith, S. C.Turner, J. R. (2019). Intracellular MLCK1 diversion reverses barrier loss to restore mucosal homeostasis. Nature Medicine, 25(4), 690-700. https://doi.org/10.1038/s41591-019-0393-7

Publisher

Springer Science and Business Media LLC

Language

en

issn

1078-8956