title: High-speed atomic force microscopy reveals strongly polarized movement of clostridial collagenase along collagen fibrils.
abstract: Bacterial collagenases involved in donor infection are widely applied in many fields due to their high activity and specificity; however, little is known regarding the mechanisms by which bacterial collagenases degrade insoluble collagen in host tissues. Using high-speed atomic force microscopy, we simultaneously visualized the hierarchical structure of collagen fibrils and the movement of a representative bacterial collagenase, Clostridium histolyticum type I collagenase (ColG), to determine the relationship between collagen structure and collagenase movement. Notably, ColG moved ~14.5 nm toward the collagen N terminus in ~3.8 s in a manner dependent on a catalytic zinc ion. While ColG was engaged, collagen molecules were not only degraded but also occasionally rearranged to thicken neighboring collagen fibrils. Importantly, we found a similarity of relationship between the enzyme-substrate interface structure and enzyme migration in collagen-collagenase and DNA-nuclease systems, which share a helical substrate structure, suggesting a common strategy in enzyme evolution.
authors.affiliation: Imaging Research Division, Bio-AFM Frontier Research Center, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Watanabe-Nakayama
authors.affiliation: Imaging Research Division, Bio-AFM Frontier Research Center, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Itami
authors.affiliation: Imaging Research Division, Bio-AFM Frontier Research Center, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kodera
authors.affiliation: Imaging Research Division, Bio-AFM Frontier Research Center, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Ando
authors.affiliation: Imaging Research Division, Bio-AFM Frontier Research Center, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Konno
mesh_terms: D047009:Hathewaya histolytica / Q000201:enzymology*; D003067:Coenzymes / Q000378:metabolism; D003094:Collagen / Q000378:metabolism*; D003012:Microbial Collagenase / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D059748:Proteolysis; D015032:Zinc / Q000378:metabolism
chemical_list: D003067:Coenzymes; D003094:Collagen; D003012:Microbial Collagenase; D015032:Zinc