The extracellular matrix (ECM) is a natural scaffold of cells in the body. It has a complex structure comprising various proteins such as collagen and hyaladherins and polysaccharides such as hyaluronan (HA). In this study, inspired by the crosslinked ECM structure, we designed a genetically engineered Link module—LinkCFQ—by fusing a microbial transglutaminase (MTG)-reactive tag to the Link module, an HA-binding domain of tumor necrosis factor-stimulated gene-6. Although the HA-specific binding property of the Link module was preserved, LinkCFQ demonstrated excellent MTG reactivity with various proteins. Furthermore, an ECM-inspired hydrogel was fabricated from an HA–gelatin mixture crosslinked via HA/Link module interaction and MTG-catalyzed isopeptide bond formation in LinkCFQ. Cell culture and mouse experiments confirmed the hydrogel’s biocompatibility and degradability. Our findings provide new insights into the design of novel biomaterials and proteins for tissue engineering, regenerative medicine, drug discovery and delivery, disease models, biofabrication, and medical devices.

Extracellular matrix-inspired hydrogel of hyaluronan-gelatin crosslinked via a link module with transglutaminase reactive sequence

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