Turning a monocovalent flavoprotein into a bicovalent flavoprotein by structure-inspired mutagenesis

Malgorzata Kopacz, Marco W Fraaije

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

A recently discovered class of bicovalent flavoproteins is an interesting group of
enzymes because of their unusual cofactor binding mode, their open active sites and the
bulky substrates they can accept. Through a sequence comparison study we have identified
a conserved sequence region in bicovalent flavoproteins that is different from monocovalent
flavoproteins. Based on this and the available structural information we have designed
mutants of the prototype monocovalent flavoprotein, 6-hydroxy-d-nicotine oxidase (6HDNO),
in order to introduce a second cofactor-protein linkage. Two amino acid replacements,
namely histidine 130 to a cysteine and leucine 138 to a histidine, were sufficient to create a
bicovalent 6HDNO. The introduced cysteine forms a covalent bond with FAD as found in
natural bicovalent flavoproteins, while the second mutation was found to be essential to …
Original languageEnglish
JournalBioorganic & Medicinal Chemistry
Volume22
Issue number20
Pages (from-to)5621-5627
ISSN0968-0896
DOIs
Publication statusPublished - 2014
Externally publishedYes

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