Molecular crowding elicits the acceleration of enzymatic crosslinking of macromolecular substrates. Issue 2 (7th November 2022)

• Record Type: Journal Article
• Authors: Sato, Ryo; Minamihata, Kosuke; Wakabayashi, Rie; Goto, Masahiro; Kamiya, Noriho
• Is Part Of: Organic & biomolecular chemistry. Volume 21:Issue 2(2023)
• Page Start: 306
• Page End: 314
• Publication Date: 2022-11-07
• Subjects: Chemistry, Organic -- Periodicals; Bioorganic chemistry -- Periodicals; Chemistry, Physical organic -- Periodicals; 547
• DOI: 10.1039/d2ob01549h
• Languages: English
• ISSNs: 1477-0520

Abstract

Abstract : Molecular crowding elicits the acceleration of enzyme-mediated protein–protein conjugation compared with that found under non-crowded conditions. Abstract : Cytoplasm contains high concentrations of biomacromolecules. Protein behavior under such crowded conditions is reportedly different from that in an aqueous buffer solution, mainly owing to the effect of volume exclusion caused by the presence of macromolecules. Using a crosslinking reaction catalyzed by microbial transglutaminase (MTG) as a model, we herein systematically determined how the substrate size affects enzymatic activity in both dilute and crowded solutions of dextran. We first observed a threefold reduction in MTG-mediated crosslinking of a pair of small peptide substrates in 15 wt% dextran solution. In contrast, when proteinaceous substrates were involved, the crosslinking rates in 15 wt% dextran solutions accelerated markedly to levels comparable with the level in the absence of dextran. Our results provide new insights into the action of enzymes with regard to macromolecular substrates under crowded conditions, of which the potential utility was demonstrated by the formation of highly crosslinked protein polymers.