Structure and Functional Properties of Bacillus subtilis Endospore Biogenesis Factor StoA

Crow, A.; Liu, Yiming; Möller, Mirja Carlsson; Brun, Nick E. Le; Hederstedt, Lars

doi:10.1074/jbc.m809566200

Cited by 14 publications

(40 citation statements)

References 44 publications

(59 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similar observations have been made for the B. subtilis thiol-disulfide oxidoreductases ResA and StoA where a water molecule bound to a Glu residue can hydrogen bond to the C-terminal buried cysteine and lower its pK a value [68], [69]. The mechanism for lowered thiol pK a values in C-x-x-C active sites involved in redox chemistry is discussed below.…”

Section: Discussionsupporting

confidence: 63%

Tuning of Thioredoxin Redox Properties by Intramolecular Hydrogen Bonds

2013

View full text Add to dashboard Cite

Thioredoxin-like proteins contain a characteristic C-x-x-C active site motif and are involved in a large number of biological processes ranging from electron transfer, cellular redox level maintenance, and regulation of cellular processes. The mechanism for deprotonation of the buried C-terminal active site cysteine in thioredoxin, necessary for dissociation of the mixed-disulfide intermediate that occurs under thiol/disulfide mediated electron transfer, is not well understood for all thioredoxin superfamily members. Here we have characterized a 8.7 kD thioredoxin (BC3987) from Bacillus cereus that unlike the typical thioredoxin appears to use the conserved Thr8 side chain near the unusual C-P-P-C active site to increase enzymatic activity by forming a hydrogen bond to the buried cysteine. Our hypothesis is based on biochemical assays and thiolate pKa titrations where the wild type and T8A mutant are compared, phylogenetic analysis of related thioredoxins, and QM/MM calculations with the BC3987 crystal structure as a precursor for modeling of reduced active sites. We suggest that our model applies to other thioredoxin subclasses with similar active site arrangements.

show abstract

Section: Discussionsupporting

confidence: 63%

Tuning of Thioredoxin Redox Properties by Intramolecular Hydrogen Bonds

2013

View full text Add to dashboard Cite

show abstract

“…Remarkably, the role of StoA in sporulation is analogous to that of ResA in cytochrome c maturation in B. subtilis . ResA and StoA are structurally very similar proteins and both are reduced by CcdA, but they have distinct and non‐overlapping substrate specificities (Crow et al. , 2009a).…”

Section: Discussionmentioning

confidence: 99%

“…We conclude that the formation and disruption of the disulphide is catalysed by BdbD and StoA respectively. The crystal structures of both these extra‐cytoplasmic thiol‐disulphide oxidoreductases have been determined (Crow et al. , 2009a,b).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Penicillin-binding protein SpoVD disulphide is a target for StoA inBacillus subtilisforespores

Liu¹,

Möller²,

Petersen

et al. 2010

Molecular Microbiology

Self Cite

View full text Add to dashboard Cite

SummaryThe bacterial endospore is a dormant and heatresistant form of life. StoA (SpoIVH) in Bacillus subtilis is a membrane-bound thioredoxin-like protein involved in endospore cortex synthesis. It is proposed to reduce disulphide bonds in hitherto unknown proteins in the intermembrane compartment of developing forespores. Starting with a bioinformatic analysis combined with mutant studies we identified the sporulation-specific, high-molecularweight, class B penicillin-binding protein SpoVD as a putative target for StoA. We then demonstrate that SpoVD is a membrane-bound protein with two exposed redox-active cysteine residues. Structural modelling of SpoVD, based on the well characterized orthologue PBP2x of Streptococcus pneumoniae, confirmed that a disulphide bond can form close to the active site of the penicillin-binding domain restricting access of enzyme substrate or functional association with other cortex biogenic proteins. Finally, by exploiting combinations of mutations in the spoVD, stoA and ccdA genes in B. subtilis cells, we present strong in vivo evidence that supports the conclusion that StoA functions to specifically break the disulphide bond in the SpoVD protein in the forespore envelope. The findings contribute to our understanding of endospore biogenesis and open a new angle to regulation of cell wall synthesis and penicillin-binding protein activity.

show abstract

“…Since FipB has two cysteines, one would have predicted only an increase of 10 kDa, but it appears that MAL-PEG significantly alters the migration of this protein on SDS-PAGE. Aberrant migration on SDS-PAGE after MAL-PEG modification has been observed by others (21). The addition of MAL-PEG without DTT produced three bands representing oxidized FipB, reduced FipB with one MAL-PEG molecule, and reduced FipB with two MAL-PEG molecules.…”

Section: Fipb Has Both Oxidoreductase and Isomerase Activities In E mentioning

confidence: 95%

FipB, an Essential Virulence Factor of Francisella tularensis subsp. tularensis, Has Dual Roles in Disulfide Bond Formation

Qin

Zhang

Clark

et al. 2014

Journal of Bacteriology

View full text Add to dashboard Cite

FipB, an essential virulence factor of Francisella tularensis, is a lipoprotein with two conserved domains that have similarity to disulfide bond formation A (DsbA) proteins and the amino-terminal dimerization domain of macrophage infectivity potentiator (Mip) proteins, which are proteins with peptidyl-prolyl cis/trans isomerase activity. This combination of conserved domains is unusual, so we further characterized the enzymatic activity and the importance of the Mip domain and lipid modification in virulence. Unlike typical DsbA proteins, which are oxidases, FipB exhibited both oxidase and isomerase activities. FipA, which also shares similarity with Mip proteins, potentiated the isomerase activity of FipB in an in vitro assay and within the bacteria, as measured by increased copper sensitivity. To determine the importance of the Mip domain and lipid modification of FipB, mutants producing FipB proteins that lacked either the Mip domain or the critical cysteine necessary for lipid modification were constructed. Both strains replicated within host cells and retained virulence in mice, though there was some attenuation. FipB formed surface-exposed dimers that were sensitive to dithiothreitol (DTT), dependent on the Mip domain and on at least one cysteine in the active site of the DsbA-like domain. However, these dimers were not essential for virulence, because the Mip deletion mutant, which failed to form dimers, was still able to replicate intracellularly and retained virulence in mice. Thus, the Mip domains of FipB and FipA impart additional isomerase functionality to FipB, but only the DsbA-like domain and oxidase activity are essential for its critical virulence functions.

show abstract

Structure and Functional Properties of Bacillus subtilis Endospore Biogenesis Factor StoA

Cited by 14 publications

References 44 publications

Tuning of Thioredoxin Redox Properties by Intramolecular Hydrogen Bonds

Tuning of Thioredoxin Redox Properties by Intramolecular Hydrogen Bonds

Penicillin-binding protein SpoVD disulphide is a target for StoA inBacillus subtilisforespores

FipB, an Essential Virulence Factor of Francisella tularensis subsp. tularensis, Has Dual Roles in Disulfide Bond Formation

Contact Info

Product

Resources

About