Lactococcus lactis TrxD represents a subgroup of thioredoxins prevalent in Gram-positive bacteria containing WCXDC active site motifs

Olof Björnberg, Petr Efler, Denis Ebong Epie, Birte Svensson, Per Hägglund

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    Abstract

    Three protein disulfide reductases of the thioredoxin superfamily from the industrially important Gram-positive Lactococcus lactis (LlTrxA, LlTrxD and LlNrdH) are compared to the "classical" thioredoxin from Escherichia coil (EcTrx1). LlTrxA resembles EcTrx1 with a WCGPC active site motif and other key residues conserved. By contrast, LlTrxD is more distantly related and contains a WCGDC motif. Bioinformatics analysis suggests that LlTrxD represents a subgroup of thioredoxins from Gram-positive bacteria. LlNrdH is a glutaredoxin-like electron donor for ribonucleotide reductase class Ib. Based on protein-protein equilibria LlTrxA (E01 = -259 mV) and LlNrdH (E01 = -238 my) show approximately 10 mV higher standard state redox potentials than the corresponding E. coil homologues, while E 01 of LlTrxD is -243 mV, more similar to glutaredoxin than "classical" thioredoxin. EcTrx1 and LlTrxA have high capacity to reduce insulin disulfides and their exposed active site thiol is alkylated at a similar rate at pH 7.0. LlTrxD on the other hand, is alkylated by iodoacetamide at almost 100 fold higher rate and shows no activity towards insulin disulfides. LlTrxA, LlTrxD and L1NrdH are all efficiently reduced by NADPH dependent thioredoxin reductase (TrxR) from L lactis and good cross-reactivity towards E. coil TrxR was observed with LITrxD as the notable exception.
    Original languageEnglish
    JournalArchives of Biochemistry and Biophysics
    Volume564
    Pages (from-to)164-172
    ISSN0003-9861
    DOIs
    Publication statusPublished - 2014

    Keywords

    • protein-protein equilibria
    • Actinomycetes and Related Organisms Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Irregular Nonsporing Gram-Positive Rods [08890] Corynebacterium genus
    • Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Endospore-forming Gram-Positives [07810] Bacillus subtilis species
    • Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Gram-Positive Cocci [07700] Lactococcus lactis species
    • Facultatively Anaerobic Gram-Negative Rods Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Enterobacteriaceae [06702] Escherichia coli species
    • Gram-Positive Cocci Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Micrococcaceae [07702] Staphylococcus aureus species
    • Mycobacteria Actinomycetes and Related Organisms Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Mycobacteriaceae [08881] Mycobacterium genus
    • electron donor
    • glutaredoxin
    • insulin disulfides
    • NADPH 53-57-6
    • ribonucleotide reductase 9068-66-0
    • thioredoxins
    • 10060, Biochemistry studies - General
    • 10062, Biochemistry studies - Nucleic acids, purines and pyrimidines
    • 10802, Enzymes - General and comparative studies: coenzymes
    • 31000, Physiology and biochemistry of bacteria
    • bioinformatics analysis mathematical and computer techniques
    • Biochemistry and Molecular Biophysics
    • BIOCHEMISTRY
    • BIOPHYSICS
    • ESCHERICHIA-COLI THIOREDOXIN
    • PROTEIN DISULFIDE-ISOMERASE
    • BACILLUS-SUBTILIS
    • BOND FORMATION
    • RIBONUCLEOTIDE REDUCTASE
    • SACCHAROMYCES-CEREVISIAE
    • DROSOPHILA-MELANOGASTER
    • GLUTATHIONE-REDUCTASE
    • OXIDIZED GLUTATHIONE
    • REDOX POTENTIALS
    • Thioredoxin
    • Lactic acid bacteria
    • Redox potential
    • Disulfide reduction
    • Thiol-disulfide exchange

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