Totally synthetic microperoxidase-11

A totally synthetic microperoxidase-11 (MP-11) is reported. Accordingly, the undecapeptide (VQKCAQCHTVE) was synthesized by solid-phase peptide synthesis followed by the thiol-ene click reaction with haemin for reconstitution. High-speed atomic force microscopy measurement conducted in water confirm...

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Bibliographic Details
Published in:	Royal Society open science Vol. 5; no. 5; p. 172311
Main Authors:	Tanabe, Junichi, Nakano, Koji, Hirata, Ryutaro, Himeno, Toshiki, Ishimatsu, Ryoichi, Imato, Toshihiko, Okabe, Hirotaka, Matsuda, Naoki
Format:	Journal Article
Language:	English
Published:	England The Royal Society Publishing 01-05-2018 The Royal Society
Subjects:	Chemistry Electrocatalytic Reaction Enzymatic Assay High-Speed Atomic Force Microscopy Microperoxidase Spectroelectrochemistry Total Synthesis spectroelectrochemistry electrocatalytic reaction high-speed atomic force microscopy enzymatic assay total synthesis microperoxidase
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Summary:	A totally synthetic microperoxidase-11 (MP-11) is reported. Accordingly, the undecapeptide (VQKCAQCHTVE) was synthesized by solid-phase peptide synthesis followed by the thiol-ene click reaction with haemin for reconstitution. High-speed atomic force microscopy measurement conducted in water confirmed the protein reconstitution by visualizing the morphological differences as animated molecular images. The synthetic MP-11 showed a considerable magnitude of catalytic activity (27%) against the natural MP-11 in the oxidation of 3,3′,5,5′-tetramethylbenzidine by hydrogen peroxide, whereas it showed very low (2.7%) activity of a synthetic variant with a point mutation (VQKCAQCMTVE, H8M). Slab waveguide spectroscopic measurements revealed that the ferrous/ferric redox reaction occurred by the direct electron transfer with specific spectral changes. Indeed, if hydrogen peroxide existed in the solution phase, the peroxidase-modified electrode showed catalytic current–voltage behaviour regardless of whether it was prepared using natural MP-11 or the synthetic MP-11. If a substrate recycling reaction was assumed, computer simulation well reproduced the experimental curves to give a global set of electrocatalytic reaction parameters. In any of the experiments, the synthetic MP-11 and natural MP-11 gave almost identical results. Our approach will be a convenient means of preparing MP-11, as well as its mutants, that does not rely on nature.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Electronic supplementary material is available online at https://dx.doi.org/10.6084/m9.figshare.c.4090226. This article has been edited by the Royal Society of Chemistry, including the commissioning, peer review process and editorial aspects up to the point of acceptance.
ISSN:	2054-5703 2054-5703
DOI:	10.1098/rsos.172311