Review
Biochemistry & Molecular Biology
Dinendra L. Abeyawardhane, Raquel Godoy-Ruiz, Kaylin A. Adipietro, Kristen M. Varney, Richard R. Rustandi, Edwin Pozharski, David J. Weber
Summary: There is currently no treatment available for toxicities caused by Clostridioides difficile binary toxin, and new therapeutic strategies are needed to address this issue.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Food Science & Technology
Katharina Ernst, Marc Landenberger, Julian Nieland, Katharina Norgaard, Manfred Frick, Giorgio Fois, Roland Benz, Holger Barth
Summary: The study investigated the cytotoxic effect of CDTb on cells, revealing that CDTb impairs cell viability and leads to redistribution of F-actin without affecting tubulin structures in CaCo-2 cells. Pharmacological pore blockers were found to effectively inhibit the cytotoxicity induced by pore formation of CDTb in the cytoplasmic membrane.
Review
Gastroenterology & Hepatology
Salvador Lopez-Cardenas, Eva Torres-Martos, Juan Mora-Delgado, Juan Manuel Sanchez-Calvo, Marta Santos-Pena, Angel Zapata Lopez, Maria Dolores Lopez-Prieto, Salvador Perez-Cortes, Juan Carlos Alados
Summary: The study revealed that patients with detected presence of TcdB and CDT in feces exhibited greater disease severity and higher recurrence rates compared to those without detection. Simultaneous detection of both markers had a stronger impact on prognosis in cases of CDI.
Review
Food Science & Technology
Adrian Martinez-Melendez, Flora Cruz-Lopez, Rayo Morfin-Otero, Hector J. Maldonado-Garza, Elvira Garza-Gonzalez
Summary: Infection with Clostridioides difficile (CDI) is a common healthcare-associated infection that causes symptoms ranging from mild diarrhea to severe pseudomembranous colitis. The binary toxin (CDT) produced by approximately 20% of C. difficile strains is thought to enhance the toxicity of toxin A and toxin B, but the role of CDT in CDI remains controversial.
Article
Immunology
Mary K. Young, Jhansi L. Leslie, Gregory R. Madden, David M. Lyerly, Robert J. Carman, Matthew W. Lyerly, David B. Stewart, Mayuresh M. Abhyankar, William A. Petri
Summary: Patients with CDT-positive fecal samples showed increased disease severity and worse clinical outcomes, with higher bacterial burden and white blood cell counts. However, there was no significant difference in gut microbiome diversity between CDT-positive and -negative patients. The detection of CDT-expressing strains may be valuable in identifying individuals at risk of severe CDI.
OPEN FORUM INFECTIOUS DISEASES
(2022)
Article
Multidisciplinary Sciences
Simoun Icho, Jennifer S. Ward, John Tam, Larry K. Kociolek, Casey M. Theriot, Roman A. Melnyk
Summary: This study investigated the impact of the total pool of intestinal bile acids on the virulence of Clostridioides difficile. It was found that the amount of bile acids in the intestine is correlated with the ability to inhibit TcdB toxicity. Even in antibiotic-treated and germ-free mice, bile acids were able to protect cells from TcdB damage, but this protection could be overcome at high toxin doses typical of severe infections. These findings reveal the role of intestinal bile acids in attenuating virulence and provide insights into asymptomatic carriage of toxigenic C. difficile, as well as strategies for manipulating bile acid levels for therapeutic benefit.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2023)
Article
Microbiology
Zohar Hamo, Maya Azrad, Boris Fichtman, Avi Peretz
Summary: Different strains of Clostridioides difficile produce varying levels of toxins that have different degrees of damage to host cells. However, no direct association was found between specific bacterial strains, toxin concentrations, and disease severity.
FRONTIERS IN MICROBIOLOGY
(2021)
Article
Microbiology
Florian Stieglitz, Ralf Gerhard, Andreas Pich
Summary: Clostridioides difficile is a major cause of nosocomial infections worldwide, mainly due to toxin A and toxin B. Hypervirulent strains producing CDT toxin are associated with higher mortality rates. The study revealed that treatment with CDTa and CDTb led to changes in phosphorylation levels of around 1100 phosphosites, indicating potential involvement of CSNK2A1 as an effector kinase for CDTa.
FRONTIERS IN MICROBIOLOGY
(2021)
Article
Immunology
Reena Thakur, Chander Raman Suri, Praveen Rishi
Summary: This study investigates the role of typhoid toxin in inducing symptoms similar to typhoid fever. The research reveals that the catalytic subunit CdtB of typhoid toxin mediates DNA damage and apoptosis of cells.
MICROBIAL PATHOGENESIS
(2022)
Article
Food Science & Technology
Aria Aminzadeh, Rene Jorgensen
Summary: By optimizing culture parameters and purification methods, the production yield of native toxin A and toxin B from Clostridioides difficile was significantly improved, resulting in high concentrations of toxin B. Additionally, optimal conditions for improving protein stability during storage were identified.
Article
Multidisciplinary Sciences
Akihiro Kawamoto, Tomohito Yamada, Toru Yoshida, Yusui Sato, Takayuki Kato, Hideaki Tsuge
Summary: Some bacteria have a toxin delivery system that transports enzymes into host cells. The bacterium Clostridioides difficile has a clinically important system with CDTa and CDTb proteins as therapeutic targets. The cryo-EM structure of CDTa bound to CDTb-pore was reported, revealing structural changes induced by CDTa binding and the involvement of an NSS-loop in substrate translocation. The dynamic information obtained provides insights for drug design against hypervirulent C. difficile strains.
NATURE COMMUNICATIONS
(2022)
Article
Microbiology
Qichao Chen, Huan Wang, Liang Li, Shibang Guo, Zhankui Liu, Zizhe Hu, Chen Tan, Huanchun Chen, Xiangru Wang
Summary: Glaesserella parasuis is the etiological agent of Glasser's disease, causing severe systemic infection. This study screened monoclonal antibodies against G. parasuis CdtB and identified one mAb, 4F10, that specifically recognizes all serotypes of G. parasuis and exhibits neutralizing activity against the cytotoxicity caused by CdtB.
VETERINARY MICROBIOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Marc Landenberger, Julian Nieland, Maurice Roeder, Katharina Norgaard, Panagiotis Papatheodorou, Katharina Ernst, Holger Barth
Summary: Clostridioides difficile produces multiple toxins, including TcdA, TcdB, and CDT, which are the main virulence factors causing associated diarrhea and colitis. CDTb alone exhibits cytotoxic effects on cells, leading to cell rounding and impairment of cell function by binding to the cell receptor LSR.
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
(2021)
Article
Immunology
Cody P. Doolan, Thomas Louie, Christopher Lata, Oscar E. Larios, William Stokes, Joseph Kim, Kristen Brown, Paul Beck, Rob Deardon, Dylan R. Pillai
Summary: A study utilized latent class analysis (LCA) to determine the genuine probability of Clostridioides difficile infection (CDI) based on laboratory and clinical variables, establishing 3 CDI probability zones. The most fitting model incorporated toxigenic culture, clinical diagnosis, and toxin antigen detection. Commercial real-time PCR platforms could delineate CDI probability zones based on Ct values obtained from LCA results.
CLINICAL INFECTIOUS DISEASES
(2021)
Article
Immunology
Morgan Simpson, Terry Bilverstone, Jhansi Leslie, Alexandra Donlan, Md Jashim Uddin, William A. Petri, Natasha Marin, Sarah Kuehne, Nigel P. Minton, William A. Petri Jr
Summary: This study found that the binding component of Clostridioides difficile binary toxin, CDTb, contributes to virulence in a hamster model of infection.
OPEN FORUM INFECTIOUS DISEASES
(2023)
Article
Allergy
Milica M. Grozdanovic, Christine B. Doyle, Li Liu, Brian T. Maybruck, Mark A. Kwatia, Nethaji Thiyagarajan, K. Ravi Acharya, Steven J. Ackerman
JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY
(2020)
Article
Biochemistry & Molecular Biology
Kyle S. Gregory, Sai Man Liu, K. Ravi Acharya
Article
Biochemistry & Molecular Biology
Gyles E. Cozier, Lizelle Lubbe, Edward D. Sturrock, K. Ravi Acharya
BIOCHEMICAL JOURNAL
(2020)
Article
Chemistry, Medicinal
Urvashi Sharma, Gyles E. Cozier, Edward D. Sturrock, K. Ravi Acharya
JOURNAL OF MEDICINAL CHEMISTRY
(2020)
Article
Biochemistry & Molecular Biology
Jonathan R. Davies, Amy Britton, Sai Man Liu, K. Ravi Acharya
Article
Biochemistry & Molecular Biology
Gyles E. Cozier, Lizelle Lubbe, Edward D. Sturrock, K. Ravi Acharya
Summary: ACE is a key enzyme involved in various physiological functions and has multiple hydrolytic activities. It contains two homologous catalytic domains (N- and C-domains) and the study of its structures provides important insights into how ACE domains can accommodate various substrates and allow different peptidase activities.
Review
Biochemistry & Molecular Biology
Samuel L. Crine, K. Ravi Acharya
Summary: C-mannosylation is a rare form of glycosylation that enhances the structural and functional diversity of proteins by forming a carbon-carbon bond with a tryptophan residue. Despite being discovered in 1994, it is still poorly understood and further research is needed to explore its mechanisms and functions.
Article
Chemistry, Medicinal
Lauren B. Arendse, Gyles E. Cozier, Charles J. Eyermann, Gregory S. Basarab, Sylva L. Schwager, Kelly Chibale, K. Ravi Acharya, Edward D. Sturrock
Summary: This study investigates the structural requirements for potent dual cACE/NEP inhibition by synthesizing and analyzing the crystal structures of inhibitors with high resolution. The results highlight the importance of the interplay between the S-1' and S-2' subsites for the selectivity of ACE domain.
JOURNAL OF MEDICINAL CHEMISTRY
(2022)
Article
Food Science & Technology
Kyle S. Gregory, Otsile O. Mojanaga, Sai Man Liu, K. Ravi Acharya
Summary: Botulinum neurotoxins (BoNT) are highly toxic proteins that cause botulism by proteolysis of a SNARE protein. They consist of three domains, with the H-C domain responsible for neuronal specificity. Some BoNTs have been commercialized for treating various neuromuscular conditions.
Article
Biochemistry & Molecular Biology
Gyles E. Cozier, Emma C. Newby, Sylva L. U. Schwager, R. Elwyn Isaac, Edward D. Sturrock, K. Ravi Acharya
Summary: This study investigates the two isoforms of human angiotensin I-converting enzyme (ACE) and their different substrate specificities and structural features. By studying the structural characteristics of each domain, more specific and potent ACE inhibitors can be designed to reduce side effects and potentially be used clinically.
Article
Food Science & Technology
Kyle S. Gregory, Tejaswini B. Mahadeva, Sai Man Liu, K. Ravi Acharya
Summary: Botulinum neurotoxins are toxic substances that can cause botulism, but they can also be used to treat neuromuscular conditions. This study reveals the crystal structures of the cell binding domain of botulinum neurotoxin A2 and its interactions with the receptor ganglioside GD1a. The findings provide important insights into the binding mechanism between the toxin and its receptor.
Article
Biochemistry & Molecular Biology
Kyle S. Gregory, Anna R. Newell, Otsile O. Mojanaga, Sai Man Liu, K. Ravi Acharya
Summary: This study determined the crystal structure of botulinum neurotoxin A6 cell binding domain (H-C/A6) in complex with GD1a and presented a new crystal form of wild type H-C/A6 (crystal form II), revealing the degree of conformational flexibility exhibited by H-C/A6.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Review
Food Science & Technology
Kyle S. Gregory, K. Ravi Acharya
Summary: Botulinum neurotoxins (BoNTs) are highly potent neurotoxins that cause neuromuscular paralysis by cleaving SNARE complex proteins. They have high affinity and specificity for neuromuscular junctions. This review focuses on the structural analysis of the cell-binding domain of different BoNT subtypes, providing insights into their functions and potential therapeutic applications.
Review
Biochemistry & Molecular Biology
Amelia Barclay, K. Ravi Acharya
Summary: Plastic pollution is a significant environmental concern, and finding enzymes capable of degrading plastics is a potential solution. However, challenges such as efficiency and scalability, as well as the diverse range of plastic waste, hinder their widespread application. Structural biology provides insights into the interactions between enzymes and plastics, which is crucial for harnessing their potential to address the plastic waste crisis.
Review
Medicine, Research & Experimental
Lizelle Lubbe, Gyles E. Cozier, Delia Oosthuizen, K. Ravi Acharya, Edward D. Sturrock
Article
Biochemical Research Methods
Juri Sakata, Toshifumi Tatsumi, Akira Sugiyama, Akihiro Shimizu, Yuya Inagaki, Hiroto Katoh, Takefumi Yamashita, Kazuki Takahashi, Sho Aki, Yudai Kaneko, Takeshi Kawamura, Mai Miura, Masazumi Ishii, Tsuyoshi Osawa, Toshiya Tanaka, Shumpei Ishikawa, Masanobu Tsukagoshi, Michael Chansler, Tatsuhiko Kodama, Motomu Kanai, Hidetoshi Tokuyama, Kenzo Yamatsugu
Summary: This study reports an expression, refolding, and purification method for antibody-mimetic drug conjugate (AMDC). The AMDC uses a variable heavy chain of heavy chain-only antibodies (VHHs) as the targeting protein and non-covalently binds to a potent DNA-alkylating drug. The AMDC shows strong cytotoxic effects on cancer cells.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Yao Wang, Shaoting Weng, Yajie Tang, Sen Lin, Xiayue Liu, Wenhui Zhang, Gang Liu, Boomi Pandi, Yinrong Wu, Lei Ma, Lin Wang
Summary: In this study, a high yield of recombinant CD20 and claudin 18.2 proteins was achieved using an in vitro coupled transcription-translation system. The results showed that rituximab has a high affinity with CD20 protein. This study provides a novel concept for promoting the expression of multi-pass transmembrane proteins and lays the foundation for large-scale industrial production of membrane-associated drug targets.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Xiaobing Chen, Zijuan Zou, Wei Li, Xu Dong, Yi Chen, Yan Lu, Mingyue Zhu, Mengsen Li, Bo Lin
Summary: In this study, the fusion protein ImI-AFP3, composed of alpha-Conotoxin ImI and human alpha fetoprotein domain 3 (AFP3), was found to inhibit the growth and migration of lung cancer cells and showed synergistic effects with the drug gefitinib. These findings suggest that ImI-AFP3 is a promising candidate for the development of anticancer drugs.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Angel Castillo-Corujo, Mirva J. Saaranen, Lloyd W. Ruddock
Summary: This study successfully expressed two Fabs antibodies in the cytoplasm of E. coli using the CyDisCo system, achieving high yields and biological activity under industrially relevant fermentation conditions.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Junling Guo, Zhongyi Cheng, Zhemin Zhou
Summary: This study focused on a halophilic archaeal nitrile hydratase (NHase) and found that it exhibited higher tolerance to substrates and products compared to NHases from other sources. The unique genetic structure of this highly stable archaeal NHase could provide a theoretical foundation for modifying and enhancing the industrial application of NHase.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Yunlong Shen, Ruirui Zhang, Xiaohua Jiang, Jinliang Yang
Summary: The study developed a monoclonal antibody, B1M023, that can bind to LILRB1 with high affinity and block its binding to HLA-G. This antibody can promote the activation and IFN-gamma secretion of T cells, suggesting its potential applications in concomitant diagnosis and tumor immunotherapy.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Jun Jiang, Zhengqiang Jiang, Qiaojuan Yan, Susu Han, Shaoqing Yang
Summary: A novel alginate lyase gene from a marine bacterium showed efficient enzyme activity and stability, producing high conversion ratios of alginate oligosaccharides.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Xianghua Xiong, Yujin Qiu, Jiahao Zheng, Ling Zhou, Qingyang Wang, Jinglun Pang, Weicai Zhang, Huipeng Chen, Gang Liu, Xiaodong Han
Summary: A specific monoclonal antibody ML419 has been found to disrupt the recognition between Botulinum neurotoxin serotype A (BoNT/A) and FGFR3, effectively preventing BoNT/A from entering neurons. In vivo experiments show that ML419 has a strong protective effect, making it a promising candidate for the development of therapeutics against BoNT/A.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Maria Rain Jennings, Soohyon Min, Grace S. Xu, Kassandra Homayuni, Bhavana Suresh, Yusef Amir Haikal, John Blazeck
Summary: This study successfully produced Homo sapiens adenosine deaminase isoform 1 (HsADA1) and its variants through optimizing the recombinant expression process. The D8N variant of HsADA1 was found to be about 30% less active than the wildtype, but it better retained its activity in human serum. Additionally, the study revealed a previously undescribed phenomenon involving albumin that contributed to the increased activity of HsADA1 and the D8N variant in serum.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Oinam Sangita Devi, Senjam Sunil Singh, K. Rana, Sorokhaibam Jibankumar Singh, Wayenbam Sobhachandra Singh
Summary: A new lectin with hemagglutination activity was purified from the rhizome of Xanthosoma violaceum Schott. The lectin showed different reactions towards human red blood cells of different blood groups. It exhibited optimal hemagglutination activity at specific temperature and pH range, and showed good stability.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
