Osmoprotection of Bacillus subtilis through Import and Proteolysis of Proline-Containing Peptides

Zaprasis, Adrienne; Brill, Jeanette; Thüring, Marietta; Wünsche, Guido; Heun, Magnus; Barzantny, Helena; Hoffmann, Tamara; Bremer, Erhard

doi:10.1128/aem.01934-12

Cited by 54 publications

(78 citation statements)

References 77 publications

(168 reference statements)

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“…The intracellular proline content of B. subtilis cells was determined by a colorimetric assay that detects L-proline as a colored proline-ninhydrin complex that can be quantified by measuring the absorption of the solution at 480 nm (45). For these assays, the B. subtilis cells were grown in SMM with 0.4 M NaCl; harvesting and processing of the cells, the details of the assay conditions, and the calculation of the intracellular volume have been described previously (15,21,53). Consistent with previous measurements (21), the proline pool of strain JH642 was found to be 95 Ϯ 12 mM in cells that had been grown in SMM containing 0.4 M NaCl; the concentrations of proline pools detected in the 15 tested DHP-resistant mutant strains ranged from 79 Ϯ 4 mM to 98 Ϯ 5 mM.…”

Section: Chemicalsmentioning

confidence: 99%

“…L-Proline serves both as a nutrient and as a stress protectant for B. subtilis since the cell can exploit it not only as a sole carbon, nitrogen, and energy source (10)(11)(12) but also as an osmostressrelieving compound (13)(14)(15)(16). To fuel protein synthesis, B. subtilis produces proline from the precursor glutamate (17,18), a pathway that is present in many bacterial species (19).…”

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confidence: 99%

“…The anabolic proline synthesis route (ProB-ProA-ProI [ProG]) is interconnected with the osmostress-relieving production route for proline (ProJ-ProA-ProH) via the ␥-glutamyl phosphate reductase (ProA) (14). The latter pathway is osmotically controlled and allows B. subtilis to produce very large amounts of the compatible solute proline (20) to fend off the detrimental effects of high salinity on cellular hydration, turgor, and physiology (13)(14)(15)21). Proline-mediated osmoprotection can also be achieved through uptake; the osmotically inducible OpuE transporter of B. subtilis serves this function (15,16).…”

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confidence: 99%

“…The latter pathway is osmotically controlled and allows B. subtilis to produce very large amounts of the compatible solute proline (20) to fend off the detrimental effects of high salinity on cellular hydration, turgor, and physiology (13)(14)(15)21). Proline-mediated osmoprotection can also be achieved through uptake; the osmotically inducible OpuE transporter of B. subtilis serves this function (15,16). OpuE also operates as a recapturing device for newly synthesized proline that is released from B. subtilis grown under high-salinity conditions, probably in an effort by the cell to fine-tune turgor (22).…”

mentioning

confidence: 99%

“…Externally provided proline not only affords osmostress protection (15,16), but it also can serve as the sole carbon, energy, and nitrogen source for B. subtilis (10)(11)(12). Utilization of proline as a nutrient requires its capture from environmental sources, such as root exudates and organic deposits in the rhizosphere (2,25), and relies on the PutB-and PutC-mediated catabolism to glutamate (10), a central intermediate in the interconnected carbon and nitrogen utilization systems of B. subtilis (26,27).…”

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confidence: 99%

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The γ-Aminobutyrate Permease GabP Serves as the Third Proline Transporter of Bacillus subtilis

et al. 2014

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bPutP and OpuE serve as proline transporters when this imino acid is used by Bacillus subtilis as a nutrient or as an osmostress protectant, respectively. The simultaneous inactivation of the PutP and OpuE systems still allows the utilization of proline as a nutrient. This growth phenotype pointed to the presence of a third proline transport system in B. subtilis. We took advantage of the sensitivity of a putP opuE double mutant to the toxic proline analog 3,4-dehydro-DL-proline (DHP) to identify this additional proline uptake system. DHP-resistant mutants were selected and found to be defective in the use of proline as a nutrient. Whole-genome resequencing of one of these strains provided the lead that the inactivation of the ␥-aminobutyrate (GABA) transporter GabP was responsible for these phenotypes. DNA sequencing of the gabP gene in 14 additionally analyzed DHPresistant strains confirmed this finding. Consistently, each of the DHP-resistant mutants was defective not only in the use of proline as a nutrient but also in the use of GABA as a nitrogen source. The same phenotype resulted from the targeted deletion of the gabP gene in a putP opuE mutant strain. Hence, the GabP carrier not only serves as an uptake system for GABA but also functions as the third proline transporter of B. subtilis. Uptake studies with radiolabeled GABA and proline confirmed this conclusion and provided information on the kinetic parameters of the GabP carrier for both of these substrates.

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Section: Chemicalsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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confidence: 99%

mentioning

confidence: 99%

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The γ-Aminobutyrate Permease GabP Serves as the Third Proline Transporter of Bacillus subtilis

et al. 2014

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Management of Osmotic Stress by Bacillus Subtilis : Genetics and Physiology

Hoffmann

Bremer

2016

Stress and Environmental Regulation of Gene Expression and Adaptation in Bacteria

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Crystal structure of a novel prolidase from Deinococcus radiodurans identifies new subfamily of bacterial prolidases

et al. 2017

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Xaa-Pro peptidases (XPP) are dinuclear peptidases of MEROPS M24B family that hydrolyze Xaa-Pro iminopeptide bond with a trans-proline at the second position of the peptide substrate. XPPs specific towards dipeptides are called prolidases while those that prefer longer oligopeptides are called aminopeptidases P. Though XPPs are strictly conserved in bacterial and archaeal species, the structural and sequence features that distinguish between prolidases and aminopeptidases P are not always clear. Here, we report 1.4 Å resolution crystal structure of a novel XPP from Deinococcus radiodurans (XPPdr). XPPdr forms a novel dimeric structure via unique dimer stabilization loops of N-terminal domains such that their C-terminal domains are placed far apart from each other. This novel dimerization is also the consequence of a different orientation of N-terminal domain in XPPdr monomer than those in other known prolidases. The enzymatic assays show that it is a prolidase with broad substrate specificity. Our structural, mutational, and molecular dynamics simulation analyses show that the conserved Arg46 of N-terminal domain is important for the dipeptide selectivity. Our BLAST search found XPPdr orthologs with conserved sequence motifs which correspond to unique structural features of XPPdr, thus identify a new subfamily of bacterial prolidases.

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Osmoprotection of Bacillus subtilis through Import and Proteolysis of Proline-Containing Peptides

Cited by 54 publications

References 77 publications

The γ-Aminobutyrate Permease GabP Serves as the Third Proline Transporter of Bacillus subtilis

The γ-Aminobutyrate Permease GabP Serves as the Third Proline Transporter of Bacillus subtilis

Management of Osmotic Stress by Bacillus Subtilis : Genetics and Physiology

Crystal structure of a novel prolidase from Deinococcus radiodurans identifies new subfamily of bacterial prolidases

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