Article
Biochemical Research Methods
Lin Yu, Nan Huang, Liangpeng Ge, Heng Sun, Yuna Fu, Chundong Liu, Jianhua Wang
Summary: Through engineering interchain disulfide bonds in the Fab region of bispecific antibody BsAb, it was found that this approach markedly increased monomeric BsAb formation and yield, improving thermostability and stability against aggregation and degradation. Additionally, the engineered BsAbs maintained their affinity and showed enhanced inhibitory effect on target cells.
JOURNAL OF BIOLOGICAL ENGINEERING
(2021)
Article
Biotechnology & Applied Microbiology
Laura Navone, Thomas Vogl, Pawarisa Luangthongkam, Jo-Anne Blinco, Carlos H. Luna-Flores, Xiaojing Chen, Juhani von Hellens, Stephen Mahler, Robert Speight
Summary: This study presents a thermostable variant of the E. coli AppA phytase, ApV1, with an extra non-consecutive disulfide bond, which shows promising thermostability but leads to decreased production in Pichia pastoris. Co-expression of folding chaperones like protein disulfide bond isomerase (Pdi) significantly increased the production of ApV1 and restored yields to levels comparable with the wild-type enzyme. Engineering for enhanced enzymatic properties may result in folding complexity and reduced production in microbial systems, highlighting the importance of developing improved production strains in parallel to achieve desirable levels of recombinant protein for industrial processes.
BIOTECHNOLOGY FOR BIOFUELS
(2021)
Review
Chemistry, Multidisciplinary
Pablo G. Argudo, Juan J. Giner-Casares
Summary: Proteins and peptide fragments play a crucial role in self-assembly for nanostructures, with intrinsically disordered proteins and protein regions showing significant biological activity. Experimental techniques and computational modeling procedures are used for characterization, leading to a wide variety of nanostructures and promising performance in biotechnological applications. Exciting possibilities for IDPs and IDRs in nanotechnology with relevant biological applications are demonstrated.
NANOSCALE ADVANCES
(2021)
Review
Biotechnology & Applied Microbiology
Marcel Passon, Stefaan De Smedt, Hristo L. Svilenov
Summary: In contrast to other species, cattle possess exceptional antibodies with ultra-long complementarity-determining regions (u1CDRs) that can consist of 40-70 amino acids. The bovine u1CDR is folded into a stalk and a disulfiderich knob domain, which enable binding to different antigens. This article summarizes the current knowledge of the u1CDR structure, discusses approaches to discover u1CDRs against novel antigens, and outlines protein engineering approaches inspired by natural u1CDRs. Furthermore, it highlights the potential of using isolated bovine knobs, known as picobodies, as small antigen-binding domains derived from natural antibodies.
BIOTECHNOLOGY ADVANCES
(2023)
Article
Biotechnology & Applied Microbiology
Laura Navone, Thomas Vogl, Pawarisa Luangthongkam, Jo-Anne Blinco, Carlos Luna-Flores, Xiaojing Chen, Juhani von Hellens, Robert Speight
Summary: The study explores potential bottlenecks in expressing E. coli AppA phytase in P. pastoris by using bidirectional promoters (BDPs) and other proteins. Transcriptional studies provide insights into the expression profile of BDPs. The use of ERV2, a flavoprotein not previously characterized in P. pastoris, improves phytase expression, suggesting it as an alternative pathway to ERO1.
MICROBIAL CELL FACTORIES
(2021)
Article
Biochemistry & Molecular Biology
Lei Zhu, Hongxin Zhao, Juanjuan Liu, Hao Cai, Bo Wu, Zhijun Liu, Shu Zhou, Qingsong Liu, Xiaokun Li, Bin Bao, Jian Liu, Han Dai, Junfeng Wang
Summary: FGF21 is a regulator of glucose and lipid metabolism, and the designed FGF21(SS) chimera shows improved thermostability and effectiveness against T2DM through structural analysis and design.
Article
Biochemistry & Molecular Biology
Xunxun Jian, Yong Wu, Zaoli Mei, Xiaopeng Zhu, Dongting Zhangsun, Sulan Luo
Summary: In the synthesis of conotoxins with multiple disulfide bonds, determining the natural disulfide bond connectivities is challenging due to the diverse connectivities formed during oxidative folding. This study focuses on KIIIA, a mu-conotoxin with potent inhibitory activity against Nav1.2 and Nav1.4. The non-natural connectivity pattern of KIIIA exhibits the highest activity. The study optimized the Fmoc solid-phase synthesis of KIIIA using different strategies, finding that random oxidation and selective strategies can produce high yields and ideal isomers. Distributed oxidation with different protecting groups was also explored, indicating the importance of cleavage order for avoiding disulfide bond scrambling. The activity of synthesized isomers on Nav1.4 was tested, providing valuable guidance for future studies on multi-disulfide-bonded peptides.
Review
Chemistry, Multidisciplinary
Bingrui Li, Peng-Fei Cao, Tomonori Saito, Alexei P. Sokolov
Summary: Self-healing materials offer new possibilities for sustainable technologies and improved device longevity. In this overview, we discuss recent developments in intrinsically self-healing polymers, which are mainly based on polymers with dynamic covalent and noncovalent bonds. We describe current self-healing mechanisms and provide examples of systems with different types of dynamic bonds. The most intriguing results are achieved when combining multiple types of dynamic bonds, resulting in materials with high toughness and fast self-healing rates. However, there is a trade-off between self-healing rate and mechanical modulus, and we propose design principles to overcome this trade-off. We also discuss applications and challenges in the field of intrinsically self-healing polymers.
Review
Biochemistry & Molecular Biology
Matthew A. H. Parson, Meredith L. Jenkins, John E. Burke
Summary: This review discusses the application of Hydrogen-deuterium exchange mass spectrometry (HDX-MS) as a powerful biophysical tool for studying the structure and function of intrinsically disordered regions in proteins. The focus is on the theory of hydrogen exchange and its practical application in identifying disordered regions and characterizing their involvement in protein-protein and protein-membrane interfaces.
BIOCHEMICAL SOCIETY TRANSACTIONS
(2022)
Review
Biochemistry & Molecular Biology
Jiahui Fu, Jihui Gao, Zhongxin Liang, Dong Yang
Summary: Disulfide bonds are crucial for maintaining protein structures and biological functions, with PDIs playing a key role in regulating their formation.
Article
Biochemistry & Molecular Biology
Shunsuke Okada, Motonori Matsusaki, Masaki Okumura, Takahiro Muraoka
Summary: This study investigated the influence of a spacer between thiol and guanidyl units on oxidative protein folding, revealing that a conjugate with a diethylene glycol spacer showed lower folding promotion effect. The reduced efficiency was likely due to the lower acidity and more reductive property of the thiol group compared to the conjugate without the spacer. This suggests that the spacer between thiol and guanidyl groups plays a critical role in promoting oxidative protein folding.
Review
Biochemistry & Molecular Biology
Sabrina L. Slater, Despoina A. I. Mavridou
Summary: Protein folding is crucial for biological function and recombinant protein production, and the correct formation of disulfide bonds is essential for protein stability. In bacterial expression systems, the limitations of endogenous posttranslational modification systems can hinder the production of proteins with their native folds.
MOLECULAR MICROBIOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Lukas A. A. Rettenbacher, Tobias von der Haar
Summary: This study establishes a novel approach for quantitatively investigating the ability of E. coli to produce disulfide bonds in its proteome. The study shows that the demand for disulfide bonded proteins in E. coli changes significantly under different growth conditions, and predicts under what conditions excess capability is available for recombinant protein production.
MICROBIAL CELL FACTORIES
(2022)
Article
Food Science & Technology
Akiko Isomoto, Eiichi Shoguchi, Kanako Hisata, Jun Inoue, Yinrui Sun, Kenji Inaba, Noriyuki Satoh, Tomohisa Ogawa, Hiroki Shibata
Summary: This study identified genes involved in the modification and functioning of venom proteins in Protobothrops flavoviridis, a venomous snake species. Genes encoding protein disulfide isomerase (PDI) family members, Selenoprotein M (SELENOM), and Calreticulin (CALR) were highly expressed in venom glands, suggesting their potential role in protein folding and modification.
Article
Biology
Motonori Matsusaki, Rina Okada, Yuya Tanikawa, Shingo Kanemura, Dai Ito, Yuxi Lin, Mai Watabe, Hiroshi Yamaguchi, Tomohide Saio, Young-Ho Lee, Kenji Inaba, Masaki Okumura
Summary: The physiological functions of proteins are determined by their unique three-dimensional structures, with conserved disulfide-catalysts and chaperone networks playing a crucial role in correct protein folding and prevention of aggregation. Disruption of these networks is implicated in pathology. Complex formation among PDIs accelerates protein folding and prevents aggregation.
Article
Multidisciplinary Sciences
Ekaterina I. Biterova, Michail N. Isupov, Ronan M. Keegan, Andrey A. Lebedev, Anil A. Sohail, Inam Liaqat, Heli I. Alanen, Lloyd W. Ruddock
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2019)
Article
Biochemistry & Molecular Biology
Anil A. Sohail, Madhuri Gaikwad, Prakash Khadka, Mirva J. Saaranen, Lloyd W. Ruddock
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2020)
Article
Biochemistry & Molecular Biology
Antti Moilanen, Lloyd W. Ruddock
JOURNAL OF BIOLOGICAL CHEMISTRY
(2020)
Article
Biochemical Research Methods
Lisette Van Tassel, Antti Moilanen, Lloyd W. Ruddock
PROTEIN EXPRESSION AND PURIFICATION
(2020)
Article
Biochemistry & Molecular Biology
Jiro Ogura, Lloyd W. Ruddock, Nariyasu Mano
FREE RADICAL BIOLOGY AND MEDICINE
(2020)
Article
Endocrinology & Metabolism
Sini Skarp, Ji-Han Xia, Qin Zhang, Marika Loija, Alice Costantini, Lloyd W. Ruddock, Outi Makitie, Gong-Hong Wei, Minna Mannikko
JOURNAL OF BONE AND MINERAL RESEARCH
(2020)
Article
Genetics & Heredity
Meredith H. Wilson, Sujith Rajan, Aidan Danoff, Richard J. White, Monica R. Hensley, Vanessa H. Quinlivan, Rosario Recacha, James H. Thierer, Frederick J. Tan, Elisabeth M. Busch-Nentwich, Lloyd Ruddock, M. Mahmood Hussain, Steven A. Farber
Article
Biochemistry & Molecular Biology
Sven T. Sowa, Antti Moilanen, Ekaterina Biterova, Mirva J. Saaranen, Lari Lehtio, Lloyd W. Ruddock
Summary: The protein pERp1 plays a crucial role in folding IgA, IgM, integrins, and maintaining ER calcium homeostasis and plasma cell mobility. Upregulated in autoimmune diseases, pERp1 is a potential therapeutic target. Belonging to the CNPY family, these ER resident saposin-like proteins are implicated in protein folding, despite lacking lipid-binding activity typically associated with saposins. The high-resolution crystal structure of human pERp1 reveals unique structural elements and sheds light on the role of CNPY proteins in ER protein folding.
JOURNAL OF MOLECULAR BIOLOGY
(2021)
Review
Biochemistry & Molecular Biology
Aatir A. Tungekar, Angel Castillo-Corujo, Lloyd W. Ruddock
Summary: Recombinant proteins are widely used in treating diseases and industrial processes, the choice of expression system depends on various factors, and Escherichia coli has been the preferred organism for recombinant protein production for over 40 years.
MICROBIAL CELL FACTORIES-BOOK
(2021)
Article
Biochemical Research Methods
Chi Zhang, Rosario Recacha, Lloyd W. Ruddock, Antti Moilanen
Summary: This study introduces an efficient method for cytoplasmic production of recFel d 1 using eukaryotic folding factors, overcoming the folding deficiencies and insoluble inclusion body formation seen in current approaches. The solubly expressed recFel d 1 is shown to contain structural disulfides, be highly stable, and sensitive to methionine sulfoxidation, with implications for functional relevance.
PROTEIN EXPRESSION AND PURIFICATION
(2021)
Article
Biochemistry & Molecular Biology
Aatir A. A. Tungekar, Lloyd W. W. Ruddock
Summary: Efficient production of therapeutic proteins in microbial hosts is crucial for accessible healthcare. The CyDisCo system has been developed for high-yield production of recombinant proteins in the cytoplasm of Escherichia coli, but redox heterogeneity remains a challenge. By mutating cysteines in the target Fc fusion proteins, we have successfully addressed the issue of redox heterogeneity.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Multidisciplinary Sciences
Aatir A. Tungekar, Rosario Recacha, Lloyd W. Ruddock
Summary: Global health challenges like the coronavirus pandemic call for an efficient system for producing diagnostic and therapeutic interventions. Antibody treatments against SARS-CoV-2 have been developed rapidly using recombinant protein production technologies and quick screening approaches. A heterologous protein production system capable of efficiently producing therapeutic antibodies against evolving pathogens is a crucial step in preparedness for future pandemics.
SCIENTIFIC REPORTS
(2023)
Article
Microbiology
Klaudia Arauzo-Aguilera, Mirva J. Saaranen, Colin Robinson, Lloyd W. Ruddock
Summary: High-value heterologous proteins with disulfide bonds are typically targeted to the periplasm via the Sec pathway, but the Tat system shows potential as an alternative because it can transport fully folded proteins. Previous studies using the TorA signal peptide, a Tat-specific signal peptide, resulted in lower yields and protein degradation. This study demonstrates that the Tat pathway, specifically through the MdoD and AmiC signal peptides, can export the disulfide bond-containing protein YebF to the periplasm and media at high levels.
Article
Multidisciplinary Sciences
Aatir A. Tungekar, Lloyd W. Ruddock
Summary: With increased accessibility and tissue penetration, smaller antibody formats such as Fab and scFv have shown potential as effective and low-cost alternatives to full-length antibodies. Microbial hosts hold promise as production hosts for antibody fragments, but production limitations arise due to low target protein yields and protein folding complexity. This study reports an alternative antibody fragment format, FabH3, designed to overcome these limitations.
SCIENTIFIC REPORTS
(2023)
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)