An Acid Up-Regulated Surface Protein of <i>Lactobacillus paracasei</i> Strain GCRL 46 is Phylogenetically Related to the Secreted Glucan- (GpbB) and Immunoglobulin-Binding (SibA) Protein of Pathogenic Streptococci
Bacterial cell wall hydrolases, including amidases and peptidases, play a critical role in peptidoglycan turnover during growth, impacting daughter cell separation, and cell death, through autolysis. When exploring the regulation of protein expression across the growth cycle of an acid-resistant str...
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MDPI AG
2019-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/20/7/1610 |
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| author | Susan J. Pepper Margaret L. Britz |
| author_facet | Susan J. Pepper Margaret L. Britz |
| author_sort | Susan J. Pepper |
| collection | DOAJ |
| description | Bacterial cell wall hydrolases, including amidases and peptidases, play a critical role in peptidoglycan turnover during growth, impacting daughter cell separation, and cell death, through autolysis. When exploring the regulation of protein expression across the growth cycle of an acid-resistant strain of <i>Lactobacillus paracasei</i>, GCRL 46, we observed temporal up-regulation of proteins in the 40–45 kDa molecular weight range for whole-cell extracts when culturing in fermenters at a controlled pH of 4.0 versus optimum growth pH of 6.3. Up-regulation of proteins in this size range was not detected in SDS-PAGE gels of the cytosolic fraction, but was routinely detected following growth at low pH in whole cells and cell debris obtained after bead beating and centrifugation, indicating a cell surface location. N-terminal sequencing and in silico analyses showed sequence similarity with proteins in the <i>L. casei</i> group (<i>L. casei</i>, <i>L. paracasei</i> and <i>L. rhamnosus</i>) which were variously annotated as uncharacterized proteins, surface antigens, possible TrsG proteins, CHAP (cysteine, histidine-dependent amidohydrolases/peptidases)-domain proteins or putative peptidoglycan <span style="font-variant: small-caps;">d</span>,<span style="font-variant: small-caps;">l</span>-endopeptidase due to the presence of a CwlO domain. This protein is a homologue of the p40 (Msp2) secreted protein of <i>L. rhamnosus</i> LGG, which is linked to probiotic functionality in this species, and is phylogenetically related to structurally-similar proteins found in <i>Enterococcus</i>, <i>Streptococcus</i> and <i>Bifidobacterium</i> species, including the glucan-binding (GbpB), surface antigen (SagA) proteins detected in pathogenic group A streptococci species as secreted, immunoglobulin-binding (SibA) proteins (also named PcsB). Three-dimensional (3D) modelling predicted structural similarities in the CHAP proteins from the <i>L. casei</i> group and streptococcal species, indicating retention of overall architecture despite sequence divergence, and an implied retention of function during evolution. A phylogenetically-related hydrolase also contained the CwlO domain with a NLPC_P60 domain, and showed similar overall but distinct architecture to the CHAP proteins. We concluded that the surface-located, CHAP protein in <i>L. casei</i> is up-regulated during long-term exposure to acidic conditions during growth but not during acid shock. |
| first_indexed | 2024-04-13T03:52:16Z |
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| id | doaj.art-6309b230ab2d4976b20ec7651935647e |
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| language | English |
| last_indexed | 2024-04-13T03:52:16Z |
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| spelling | doaj.art-6309b230ab2d4976b20ec7651935647e2022-12-22T03:03:46ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-03-01207161010.3390/ijms20071610ijms20071610An Acid Up-Regulated Surface Protein of <i>Lactobacillus paracasei</i> Strain GCRL 46 is Phylogenetically Related to the Secreted Glucan- (GpbB) and Immunoglobulin-Binding (SibA) Protein of Pathogenic StreptococciSusan J. Pepper0Margaret L. Britz1Monash Sustainable Development Institute, 8 Scenic Boulevard, Monash University, Clayton, VIC 3800, AustraliaCentre for Food Safety and Innovation, Tasmanian Institute of Agriculture, Life Sciences Building, University of Tasmania, Hobart, TAS 7005, AustraliaBacterial cell wall hydrolases, including amidases and peptidases, play a critical role in peptidoglycan turnover during growth, impacting daughter cell separation, and cell death, through autolysis. When exploring the regulation of protein expression across the growth cycle of an acid-resistant strain of <i>Lactobacillus paracasei</i>, GCRL 46, we observed temporal up-regulation of proteins in the 40–45 kDa molecular weight range for whole-cell extracts when culturing in fermenters at a controlled pH of 4.0 versus optimum growth pH of 6.3. Up-regulation of proteins in this size range was not detected in SDS-PAGE gels of the cytosolic fraction, but was routinely detected following growth at low pH in whole cells and cell debris obtained after bead beating and centrifugation, indicating a cell surface location. N-terminal sequencing and in silico analyses showed sequence similarity with proteins in the <i>L. casei</i> group (<i>L. casei</i>, <i>L. paracasei</i> and <i>L. rhamnosus</i>) which were variously annotated as uncharacterized proteins, surface antigens, possible TrsG proteins, CHAP (cysteine, histidine-dependent amidohydrolases/peptidases)-domain proteins or putative peptidoglycan <span style="font-variant: small-caps;">d</span>,<span style="font-variant: small-caps;">l</span>-endopeptidase due to the presence of a CwlO domain. This protein is a homologue of the p40 (Msp2) secreted protein of <i>L. rhamnosus</i> LGG, which is linked to probiotic functionality in this species, and is phylogenetically related to structurally-similar proteins found in <i>Enterococcus</i>, <i>Streptococcus</i> and <i>Bifidobacterium</i> species, including the glucan-binding (GbpB), surface antigen (SagA) proteins detected in pathogenic group A streptococci species as secreted, immunoglobulin-binding (SibA) proteins (also named PcsB). Three-dimensional (3D) modelling predicted structural similarities in the CHAP proteins from the <i>L. casei</i> group and streptococcal species, indicating retention of overall architecture despite sequence divergence, and an implied retention of function during evolution. A phylogenetically-related hydrolase also contained the CwlO domain with a NLPC_P60 domain, and showed similar overall but distinct architecture to the CHAP proteins. We concluded that the surface-located, CHAP protein in <i>L. casei</i> is up-regulated during long-term exposure to acidic conditions during growth but not during acid shock.https://www.mdpi.com/1422-0067/20/7/1610<i>Lactobacillus casei</i><i>Streptococcus</i>SibACHAPPcsBcell-wall hydrolaseacid resistance |
| spellingShingle | Susan J. Pepper Margaret L. Britz An Acid Up-Regulated Surface Protein of <i>Lactobacillus paracasei</i> Strain GCRL 46 is Phylogenetically Related to the Secreted Glucan- (GpbB) and Immunoglobulin-Binding (SibA) Protein of Pathogenic Streptococci International Journal of Molecular Sciences <i>Lactobacillus casei</i> <i>Streptococcus</i> SibA CHAP PcsB cell-wall hydrolase acid resistance |
| title | An Acid Up-Regulated Surface Protein of <i>Lactobacillus paracasei</i> Strain GCRL 46 is Phylogenetically Related to the Secreted Glucan- (GpbB) and Immunoglobulin-Binding (SibA) Protein of Pathogenic Streptococci |
| title_full | An Acid Up-Regulated Surface Protein of <i>Lactobacillus paracasei</i> Strain GCRL 46 is Phylogenetically Related to the Secreted Glucan- (GpbB) and Immunoglobulin-Binding (SibA) Protein of Pathogenic Streptococci |
| title_fullStr | An Acid Up-Regulated Surface Protein of <i>Lactobacillus paracasei</i> Strain GCRL 46 is Phylogenetically Related to the Secreted Glucan- (GpbB) and Immunoglobulin-Binding (SibA) Protein of Pathogenic Streptococci |
| title_full_unstemmed | An Acid Up-Regulated Surface Protein of <i>Lactobacillus paracasei</i> Strain GCRL 46 is Phylogenetically Related to the Secreted Glucan- (GpbB) and Immunoglobulin-Binding (SibA) Protein of Pathogenic Streptococci |
| title_short | An Acid Up-Regulated Surface Protein of <i>Lactobacillus paracasei</i> Strain GCRL 46 is Phylogenetically Related to the Secreted Glucan- (GpbB) and Immunoglobulin-Binding (SibA) Protein of Pathogenic Streptococci |
| title_sort | acid up regulated surface protein of i lactobacillus paracasei i strain gcrl 46 is phylogenetically related to the secreted glucan gpbb and immunoglobulin binding siba protein of pathogenic streptococci |
| topic | <i>Lactobacillus casei</i> <i>Streptococcus</i> SibA CHAP PcsB cell-wall hydrolase acid resistance |
| url | https://www.mdpi.com/1422-0067/20/7/1610 |
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