A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Semisynthetic Nanoreactor for Reversible Single-Molecule Covalent Chemistry
Tekijät: Lee J, Boersma AJ, Boudreau MA, Cheley S, Daltrop O, Li JW, Tamagaki H, Bayley H
Kustantaja: AMER CHEMICAL SOC
Julkaisuvuosi: 2016
Journal: ACS Nano
Tietokannassa oleva lehden nimi: ACS NANO
Lehden akronyymi: ACS NANO
Vuosikerta: 10
Numero: 9
Aloitussivu: 8843
Lopetussivu: 8850
Sivujen määrä: 8
ISSN: 1936-0851
DOI: https://doi.org/10.1021/acsnano.6b04663
Tiivistelmä
Protein engineering has been used to remodel pores for applications in biotechnology. For example, the heptameric alpha-hemolysin pore (alpha HL) has been engineered to form a nanoreactor to study covalent chemistry at the single -molecule level. Previous work has been confined largely to the chemistry of cysteine side chains or, in one instance, to an irreversible reaction of an unnatural amino acid side chain bearing a terminal alkyne. Here, we present four different alpha HL pores obtained by coupling either two or three fragments by native chemical ligation (NCL). The synthetic alpha HL monomers were folded and incorporated into heptameric pores. The functionality of the pores was validated by hemolysis assays and by single-channel current recording. By using NCL to introduce a ketone amino acid, the nanoreactor approach was extended to an investigation of reversible covalent chemistry on an unnatural side chain at the single -molecule level.
Protein engineering has been used to remodel pores for applications in biotechnology. For example, the heptameric alpha-hemolysin pore (alpha HL) has been engineered to form a nanoreactor to study covalent chemistry at the single -molecule level. Previous work has been confined largely to the chemistry of cysteine side chains or, in one instance, to an irreversible reaction of an unnatural amino acid side chain bearing a terminal alkyne. Here, we present four different alpha HL pores obtained by coupling either two or three fragments by native chemical ligation (NCL). The synthetic alpha HL monomers were folded and incorporated into heptameric pores. The functionality of the pores was validated by hemolysis assays and by single-channel current recording. By using NCL to introduce a ketone amino acid, the nanoreactor approach was extended to an investigation of reversible covalent chemistry on an unnatural side chain at the single -molecule level.
Ladattava julkaisu This is an electronic reprint of the original article. |  |
Turun yliopiston Tutkimustietojärjestelmän portaali