Crystal structure of a Burkholderia peptidase and modification of the substrate-binding site for enhanced hydrolytic activity toward gluten-derived pro-immunogenic peptides

Liu, Yu-You; Lin, I-Chen; Chen, Pei-Cih; Lee, Cheng-Cheng; Meng, Menghsiao

doi:10.1016/j.ijbiomac.2022.10.016

Cited by 4 publications

(7 citation statements)

References 37 publications

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“…The crystal structure of Bga1903, highlighting residues constituting the S1–S3 subsites ( Figure 1 ), indicates that the broader specificity for P1 residues may stem from the larger cavity of S1 compared to S2 and S3. An earlier study investigated the importance of residues E380 and S387, which line the wall of S1, in influencing the peptidase’s catalytic efficiency toward specific chromogenic peptidyl substrates [ 22 ]. The present study conducts a more detailed examination of these two residues, anticipating that tailoring S1 will result in changes in selectivity for acceptable P1 residues.…”

Section: Resultsmentioning

confidence: 99%

“…Bga1903 has the capability of being secreted into the culture medium by Escherichia coli BL21(DE3). The structure of Bga1903 was determined through X-ray crystallography, and the architecture of the substrate-binding subsites was defined [ 22 ]. In the present study, an endeavor was made to further enhance its hydrolytic activity toward the 33- and 26-mer peptides by rationally modifying the S1 subsite of Bga1903 through mutagenesis.…”

Section: Introductionmentioning

confidence: 99%

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Specificity Enhancement of Glutenase Bga1903 toward Celiac Disease-Eliciting Pro-Immunogenic Peptides via Active-Site Modification

Liu,

Ye,

Meng

2023

IJMS

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Celiac disease is an autoimmune disease triggered by oral ingestion of gluten, with certain gluten residues resistant to digestive tract enzymes. Within the duodenum, the remaining peptides incite immunogenic responses, including the generation of autoantibodies and inflammation, leading to irreversible damage. Our previous exploration unveiled a glutenase called Bga1903 derived from the Gram-negative bacterium Burkholderia gladioli. The cleavage pattern of Bga1903 indicates its moderate ability to mitigate the toxicity of pro-immunogenic peptides. The crystal structure of Bga1903, along with the identification of subsites within its active site, was determined. To improve its substrate specificity toward prevalent motifs like QPQ within gluten peptides, the active site of Bga1903 underwent site-directed mutagenesis according to structural insights and enzymatic kinetics. Among the double-site mutants, E380Q/S387L exhibits an approximately 34-fold increase in its specificity constant toward the QPQ sequence, favoring glutamines at the P1 and P3 positions compared to the wild type. The increased specificity of E380Q/S387L not only enhances its ability to break down pro-immunogenic peptides but also positions this enzyme variant as a promising candidate for oral therapy for celiac disease.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Specificity Enhancement of Glutenase Bga1903 toward Celiac Disease-Eliciting Pro-Immunogenic Peptides via Active-Site Modification

Liu,

Ye,

Meng

2023

IJMS

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“…Studies propose that CD may interfere with the normal mineralization of dental enamel, resulting in structural abnormalities [ 16 ]. Enamel defects, such as pitting, grooving, and discoloration, have been reported in individuals with CD, providing potential oral markers for the condition [ 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 ].…”

Section: Introductionmentioning

confidence: 99%

Celiac Disease-Related Enamel Defects: A Systematic Review

Inchingolo,

Dipalma,

Viapiano

et al. 2024

JCM

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Introduction: This systematic review aims to elucidate the intricate correlation between celiac disease (CD) and dental enamel defects (DED), exploring pathophysiological mechanisms, oral health implications, and a dentist’s role in early diagnosis. Materials and methods: Following PRISMA guidelines, a comprehensive search from 1 January 2013 to 1 January 2024 across PubMed, Cochrane Library, Scopus, and Web of Science identified 153 publications. After exclusions, 18 studies met the inclusion criteria for qualitative analysis. Inclusion criteria involved study types (RCTs, RCCTs, case series), human participants, English language, and full-text available. Results: The search yielded 153 publications, with 18 studies meeting the inclusion criteria for qualitative analysis. Notable findings include a high prevalence of DED in CD patients, ranging from 50 to 94.1%. Symmetrical and chronological defects, according to Aine’s classification, were predominant, and significant associations were observed between CD severity and enamel defect extent. Conclusions: The early recognition of oral lesions, particularly through Aine’s classification, may signal potential CD even in the absence of gastrointestinal symptoms. Correlations between CD and dental health conditions like molar incisor hypomineralization (MIH) emphasize the dentist’s crucial role in early diagnosis. Collaboration between dentists and gastroenterologists is essential for effective monitoring and management. This review consolidates current knowledge, laying the groundwork for future research and promoting interdisciplinary collaboration for improved CD-related oral health outcomes. Further large-scale prospective research is recommended to deepen our understanding of these issues.

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“…Bga1903 belongs to the S8 peptidase family according to MEROPS classification and exhibits notable activity in breaking down the 33- and 26-mer peptides, with a particular preference for glutamine located at the P1 position. Active-site modifications based on the crystal structure of Bga1903 further enhanced the proteolytic activity of Bga1903 toward GIPs . It is known that the gastric pH after a meal could decrease from pH 6.0 down to 2.0 in about 2 h .…”

Section: Introductionmentioning

confidence: 99%

“…Active-site modifications based on the crystal structure of Bga1903 further enhanced the proteolytic activity of Bga1903 toward GIPs. 27 It is known that the gastric pH after a meal could decrease from pH 6.0 down to 2.0 in about 2 h. 28 Therefore, the insignificant activity of Bga1903 at pH < 3.5 may compromise its potential value in helping CeD patients. Screening for microorganisms capable of secreting acidic peptidases was thus continued using agar plates at pH 3.0.…”

Section: ■ Introductionmentioning

confidence: 99%

Hydrolysis of Gluten-Derived Celiac Disease-Triggering Peptides across a Broad pH Range by RmuAP1: A Novel Aspartic Peptidase Isolated from Rhodotorula mucilaginosa

Zhang,

Leu,

Meng

2023

J. Agric. Food Chem.

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An aspartate peptidase with proteolytic activity toward gluten was identified from an isolated red yeast Rhodotorula mucilaginosa strain. This peptidase consists of 425 amino acids, comprising an N-terminal signal peptide, a propeptide, and a C-terminal catalytic domain. The catalytic domain, termed RmuAP1CD, could be secreted by the recombinant oleaginous yeast Yarrowia lipolytica, whose genome contains the expression cassette for RmuAP1CD. RmuAP1CD exhibited optimum activity at pH 2.5 when acting on bovine serum albumin. Moreover, it facilitated the hydrolysis of gluten-derived immunogenic peptides (GIPs), which are responsible for triggering celiac disease symptoms, across a pH range of 3.0–6.0. The preferred cleavage sites are P–Q–Q–↓–P–Q in the 26-mer and P–Q–L–↓–P–Y in the 33-mer GIPs. Conversely, porcine pepsin cannot hydrolyze these two GIPs. The ability of RmuAP1CD to degrade GIPs under acidic conditions of the stomach indicates its potential as a viable oral enzyme therapy for celiac disease.

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Crystal structure of a Burkholderia peptidase and modification of the substrate-binding site for enhanced hydrolytic activity toward gluten-derived pro-immunogenic peptides

Cited by 4 publications

References 37 publications

Specificity Enhancement of Glutenase Bga1903 toward Celiac Disease-Eliciting Pro-Immunogenic Peptides via Active-Site Modification

Specificity Enhancement of Glutenase Bga1903 toward Celiac Disease-Eliciting Pro-Immunogenic Peptides via Active-Site Modification

Celiac Disease-Related Enamel Defects: A Systematic Review

Hydrolysis of Gluten-Derived Celiac Disease-Triggering Peptides across a Broad pH Range by RmuAP1: A Novel Aspartic Peptidase Isolated from Rhodotorula mucilaginosa

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