Intracellular MLCK1 diversion reverses barrier loss to restore mucosal homeostasis.

Abstract

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.

• Authors: W Vallen Graham, Weiqi He, Amanda M Marchiando, Juanmin Zha, Gurminder Singh, Hua-Shan Li, Amlan Biswas, Ma Lora Drizella M Ong, Zhi-Hui Jiang, Wangsun Choi, Harmon Zuccola, Yitang Wang, James Griffith, Jingshing Wu, Harry J Rosenberg, Yingmin Wang, Scott B Snapper, David Ostrov, Stephen C Meredith, Lawrence W Miller, Jerrold R Turner
• Journal: Nature medicine (Nat Med) Vol. 25 Issue 4 Pg. 690-700 (04 2019) ISSN: 1546-170X [Electronic] United States
• PMID: 30936544 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Myosin Light Chains
• Small Molecule Libraries
• Tumor Necrosis Factor-alpha
• Actomyosin
• Myosin-Light-Chain Kinase

Topics

• Actomyosin (metabolism)
• Animals
• Caco-2 Cells
• Chronic Disease
• Homeostasis (drug effects)
• Humans
• Inflammation (pathology)
• Inflammatory Bowel Diseases (pathology)
• Intestinal Mucosa (drug effects, metabolism)
• Intracellular Space (enzymology)
• Jejunum (drug effects, metabolism, pathology)
• Mice
• Myosin Light Chains (metabolism)
• Myosin-Light-Chain Kinase (chemistry, metabolism)
• Phosphorylation (drug effects)
• Protein Domains
• Small Molecule Libraries (pharmacology)
• Tight Junctions (drug effects, metabolism)
• Tumor Necrosis Factor-alpha (pharmacology)