Journal article
Lactococcus lactis TrxD represents a subgroup of thioredoxins prevalent in Gram-positive bacteria containing WCXDC active site motifs
Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark1
Department of Systems Biology, Technical University of Denmark2
Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark3
Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark4
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.
| Language: | English |
|---|---|
| Year: | 2014 |
| Pages: | 164-172 |
| ISSN: | 10960384 , 00039861 and 05706963 |
| Types: | Journal article |
| DOI: | 10.1016/j.abb.2014.09.010 |
| ORCIDs: | Svensson, Birte |
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 Actinomycetes and Related Organisms Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Irregular Nonsporing Gram-Positive Rods [08890] Corynebacterium genus BACILLUS-SUBTILIS BIOCHEMISTRY BIOPHYSICS BOND FORMATION Biochemistry and Molecular Biophysics DROSOPHILA-MELANOGASTER Disulfide reduction ESCHERICHIA-COLI THIOREDOXIN 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 GLUTATHIONE-REDUCTASE Gram-Positive Cocci Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Micrococcaceae [07702] Staphylococcus aureus species Lactic acid bacteria Mycobacteria Actinomycetes and Related Organisms Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Mycobacteriaceae [08881] Mycobacterium genus NADPH 53-57-6 OXIDIZED GLUTATHIONE PROTEIN DISULFIDE-ISOMERASE REDOX POTENTIALS RIBONUCLEOTIDE REDUCTASE Redox potential SACCHAROMYCES-CEREVISIAE Thiol-disulfide exchange Thioredoxin bioinformatics analysis mathematical and computer techniques electron donor glutaredoxin insulin disulfides protein-protein equilibria ribonucleotide reductase 9068-66-0 thioredoxins
