Exploring lectin-like activity of the S-layer protein of Lactobacillus acidophilus ATCC 4356

Martin, Joaquina Fina; Palomino, María Mercedes; Cutine, Anabella M; Modenutti, Carlos P.; Porto, Darío Augusto Fernández Do; Allievi, Mariana C.; Zanini, Sofia H; Mariño, Karina; Barquero, Andrea Alejandra; Ruzal, Sandra M.

doi:10.1007/s00253-019-09795-y

Cited by 25 publications

(54 citation statements)

References 74 publications

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“…Lactobacillus acidophilus Slps are known to anchor to the cell exterior through interactions with lipoteichoic acid (LTA), a surface-associated adhesion amphiphile (Antikainen et al, 2002;Fina Martin et al, 2019). Increased Slp shedding by igdA suggests not only a function in S-layer formation, but also a probable relationship with LTA, though notably, ltaS expression was unaffected by the deletion.…”

Section: Discussionmentioning

confidence: 99%

Deletion of S-Layer Associated Ig-Like Domain Protein Disrupts the Lactobacillus acidophilus Cell Surface

et al. 2020

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Bacterial surface-layers (S-layers) are crystalline arrays of repeating proteinaceous subunits that coat the exterior of many cell envelopes. S-layers have demonstrated diverse functions in growth and survival, maintenance of cell integrity, and mediation of host interactions. Additionally, S-layers can act as scaffolds for the outward display of auxiliary proteins and glycoproteins. These non-covalently bound S-layer associated proteins (SLAPs) have characterized roles in cell division, adherence to intestinal cells, and modulation of the host immune response. Recently, IgdA (LBA0695), a Lactobacillus acidophilus SLAP that possesses a Group 3 immunoglobulin (Ig)-like domain and GW (Gly-Tryp) dipeptide surface anchor, was recognized for its high conservation among S-layer-forming lactobacilli, constitutive expression, and surface localization. These findings prompted its selection for examination within the present study. Although IgdA and corresponding orthologs were shown to be unique to host-adapted lactobacilli, the Ig domain itself was specific to vertebrate-adapted species suggesting a role in vertebrate adaptation. Using a counterselective gene replacement system, igdA was deleted from the L. acidophilus NCFM chromosome. The resultant mutant, NCK2532, exhibited a visibly disrupted cell surface which likely contributed to its higher salt sensitivity, severely reduced adhesive capacity, and altered immunogenicity profile. Transcriptomic analyses revealed the induction of several stress response genes and secondary surface proteins. Due to the broad impact of IgdA on the cellular physiology and probiotic attributes of L. acidophilus, identification of similar proteins in alternative bacterial species may help pinpoint next-generation host-adapted probiotic candidates.

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Section: Discussionmentioning

confidence: 99%

Deletion of S-Layer Associated Ig-Like Domain Protein Disrupts the Lactobacillus acidophilus Cell Surface

et al. 2020

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“…As shown in Figure 1A, two copies of a 60-amino acids protein domain denominated SLP-A (pfam03217; Figure 1A; Boot et al, 1995) were identified within de carboxy-terminal of SlpA. Both copies of the SLP-A domain (dSLP-A) are required for an efficient cell wall binding of SlpA to the surface of L. acidophilus (Fina Martin et al, 2019). In addition, as shown in Figure 1A, the amino-terminal of SlpA (SlpA 1-31 ) encodes a predicted signal peptide (analyzed by a deep neural networkbased method SignalP-5.0) and the peptide region SlpA 31-284 for a Herpes BLLF1 protein domain (pfam05109), a protein region that has been shown to be required for self-assembly of SlpA on the bacterial surface to form the S-layer (Smit et al, 2002).…”

Section: Proteins Fused In Frame With the Dslp-a Protein Domain Can Amentioning

confidence: 99%

“…It has been described that the S-layer can be removed from the bacterial surface by cation substitution solutions (LiCl) or hydrogen bondbreaking agents such as guanidine hydrochloride (GHCl) or urea (Sleytr et al, 2014). Interestingly, after dialysis, the isolated S-layer protein can associate back to the Lactobacillus cell wall or adhere to inert surfaces that can be adapted for biotechnological purposes (Sleytr et al, 2014;Fina Martin et al, 2019). SlpA encodes in its carboxy-terminal (SlpA 284-444 ) a tandem of two copies of the protein domain denominated SLP-A (dSLP-A), a region that was shown to be necessary and sufficient for the association of SlpA to the bacterial cell wall (Fina Martin et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

“…Interestingly, after dialysis, the isolated S-layer protein can associate back to the Lactobacillus cell wall or adhere to inert surfaces that can be adapted for biotechnological purposes (Sleytr et al, 2014;Fina Martin et al, 2019). SlpA encodes in its carboxy-terminal (SlpA 284-444 ) a tandem of two copies of the protein domain denominated SLP-A (dSLP-A), a region that was shown to be necessary and sufficient for the association of SlpA to the bacterial cell wall (Fina Martin et al, 2019). Teichoic acid and wall teichoic acid present on the lactobacilli bacterial cell wall have been characterized as natural ligands for the dSLP-A domain (Fina Martin et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

“…SlpA encodes in its carboxy-terminal (SlpA 284-444 ) a tandem of two copies of the protein domain denominated SLP-A (dSLP-A), a region that was shown to be necessary and sufficient for the association of SlpA to the bacterial cell wall (Fina Martin et al, 2019). Teichoic acid and wall teichoic acid present on the lactobacilli bacterial cell wall have been characterized as natural ligands for the dSLP-A domain (Fina Martin et al, 2019). The dSLP-A domain (pfam: 03217) is found only in Lactobacillus species, limited to S-layer forming lactobacilli of the L. acidophilus group, including Lactobacillus helveticus, Lactobacillus crispatus, Lactobacillus gallinarum, and Lactobacillus amylovorus (Avall-Jaaskelainen and Palva, 2005).…”

Section: Introductionmentioning

confidence: 99%

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Development of an Antigen Delivery Platform Using Lactobacillus acidophilus Decorated With Heterologous Proteins: A Sheep in Wolf’s Clothing Story

et al. 2020

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S-layers are bacterial structures present on the surface of several Gram-positive and Gram-negative bacteria that play a role in bacterial protection. In Lactobacillus acidophilus (L. acidophilus ATCC 4356), the S-layer is mainly composed of the protein SlpA. A tandem of two copies of the protein domain SLP-A (pfam: 03217) was identified at the C-terminal of SlpA, being this double SLP-A protein domain (in short dSLP-A) necessary and sufficient for the association of the protein to the L. acidophilus cell wall. A variety of proteins fused to the dSLP-A domain were able to spontaneously associate with high affinity to the cell wall of L. acidophilus and Bacillus subtilis var. natto, in a process that we termed decoration. Binding of dSLP-A-containing-proteins to L. acidophilus was stable at conditions that mimic the gastrointestinal transit in terms of pH, proteases, and bile salts. To evaluate if protein decoration of L. acidophilus can be adapted to generate an oral vaccine platform, a chimeric antigen derived from the bacterial pathogen Shiga-toxin-producing Escherichia coli (STEC) was constructed by fusing the sequences encoding the polypeptides EspA 36-192 , Intimin 653-953 , Tir 240-378 , and H7 flagellin 352-374 (EITH7) to the dSLP-A domain (EITH7-dSLP-A). Recombinantly expressed EITH7-dSLP-A protein was affinity purified and combined with L. acidophilus cultures to allow the association of the chimeric antigen to the bacterial surface. EITH7-decorated L. acidophilus was orally administered to BALB/c mice and the induction of anti-EITH7 specific antibodies in sera and feces determined by ELISA. Mice presenting significantly higher anti-EITH7 antibodies titers were able to control more efficiently an experimental STEC infection than mice that received the non-decorated L. acidophilus carrier, indicating that antigen-decorated L. acidophilus can be adapted as a mucosal immunization delivery platform to elicit a protective immune response for vaccine purposes.

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Strategies to display heterologous proteins on the cell surface of lactic acid bacteria using as anchor the C-terminal domain of Lactobacillus acidophilus SlpA

Gordillo

Palumbo

Allievi

et al. 2020

World J Microbiol Biotechnol

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The surface-layer (S-layer) protein of Lactobacillus acidophilus is a crystalline array of self-assembling subunits, noncovalently bound to the most outer cell wall envelope, which constitutes up to 20% of the total cell protein content. These attributes make S-layer proteins an excellent anchor for the development of microbial cell-surface display systems. In L. acidophilus, the S-layer is formed predominantly by the protein SlpA. We have previously shown that the C-terminal domain of SlpA is responsible for the cell wall anchorage on L. acidophilus ATCC 4356. In the present study, we evaluated the C-terminal domain of SlpA of L. acidophilus ATCC 4356 as a potential anchor domain to display functional proteins on the surface of non-genetically modified lactic acid bacteria (LAB). To this end, green fluorescent protein (GFP)-CTSlpA was firstly produced in Escherichia coli and the recombinant proteins were able to spontaneously bind to the cell wall of LAB in a binding assay. GFP was successfully displayed on the S-layer stripped surface of L. acidophilus. Both the binding stability and cell survival of L. acidophilus decorated with the recombinant protein were then studied in simulated gastrointestinal conditions. Furthermore, NaCl was tested as a safer alternative to LiCl for S-layer removal. This study presents the development of a protein delivery platform involving L. acidophilus, a microorganism generally regarded as safe, which utilizes the contiguous, non-covalently attached S-layer at the cell surface of the bacterium without introducing any genetic modification.

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Exploring lectin-like activity of the S-layer protein of Lactobacillus acidophilus ATCC 4356

Cited by 25 publications

References 74 publications

Deletion of S-Layer Associated Ig-Like Domain Protein Disrupts the Lactobacillus acidophilus Cell Surface

Deletion of S-Layer Associated Ig-Like Domain Protein Disrupts the Lactobacillus acidophilus Cell Surface

Development of an Antigen Delivery Platform Using Lactobacillus acidophilus Decorated With Heterologous Proteins: A Sheep in Wolf’s Clothing Story

Strategies to display heterologous proteins on the cell surface of lactic acid bacteria using as anchor the C-terminal domain of Lactobacillus acidophilus SlpA

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