Article
Chemistry, Multidisciplinary
Martin D. Gelenter, Ad Bax
Summary: Post-translational modifications are commonly found in eukaryotic proteomes. However, these modifications are rarely studied using NMR in eukaryotic proteins, which are usually produced in E. coli through recombinant expression. The native C-terminal amide of melittin, a major peptide in honey bee venom, significantly affects its structure and dynamics. This study presents a method to produce isotopically labeled native melittin through recombinant expression and chemical amidation. It also confirms the accuracy of structural models generated by AlphaFold-Multimer using experimental residual dipolar couplings.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2023)
Article
Biology
Samsuzzoha Mondal, Honey Priya James, Francesco Milano, Rui Jin, Tobias Baumgart
Summary: BAR proteins, such as Amphiphysin, are known for generating membrane curvature during endocytosis. We successfully expressed and purified recombinant Amphiphysin and its N-BAR domain using affinity chromatography and GST tags. The cleavage of the GST tag in the N-BAR domain caused precipitation, which was minimized by adding glycerol to the purification buffers. Size exclusion chromatography was used to remove any oligomeric species. This protocol can also be applied to purify other N-BAR proteins.
Article
Cell Biology
Fotis L. Kyrilis, Dmitry A. Semchonok, Ioannis Skalidis, Christian Tuting, Farzad Hamdi, Francis J. O'Reilly, Juri Rappsilber, Panagiotis L. Kastritis
Summary: The study successfully identified fully assembled pyruvate dehydrogenase complex in native cell extracts and characterized its structure using multiple techniques. The findings provide a framework for further understanding the structure and function of the complex.
Article
Biochemistry & Molecular Biology
Madhuri Thool, Chandrima Dey, Srirupa Bhattacharyya, S. Sudhagar, Rajkumar P. Thummer
Summary: SOX2 transcription factor plays a critical role in early embryonic development and maintenance of stem cell identity. The study successfully expressed and purified human SOX2 protein in soluble form from a bacterial system, demonstrating its potential for integration-free stem cell derivation. This molecular tool provides opportunities for investigating SOX2's role in various cellular processes and diseases, as well as facilitating structural and biochemical studies.
MOLECULAR BIOTECHNOLOGY
(2021)
Article
Materials Science, Paper & Wood
Larissa Bussler, Debora Jacomini, Juliana Moco Correa, Marina Kimiko Kadowaki, Alexandre Maller, Rita de Cassia Garcia Simao
Summary: The cellulase from Caulobacter crescentus (C. crescentus) showed higher expression in the presence of corn straw and released more reducing sugar during the hydrolysis of corn straw and corn cob, suggesting its potential as an alternative for the production of chemicals and biofuels. Additionally, the application of the enzyme on denim jeans resulted in a cleaner and smoother surface of the fibers, indicating its use for biopolishing. This study supports the biotechnological capabilities of the cellulase from C. crescentus based on the bacterial genomic sequence.
Article
Biotechnology & Applied Microbiology
Krishna Kumar Haridhasapavalan, Pradeep Kumar Sundaravadivelu, Srirupa Bhattacharyya, Sujal Harsh Ranjan, Khyati Raina, Rajkumar P. Thummer
Summary: GATA4 transcription factor plays a vital role in early embryonic development, cardiomyocyte derivation, and tumor suppression. By optimizing expression parameters and purifying the protein, it has been confirmed that the recombinant GATA4 is non-toxic, can enter cells, and translocate to the nucleus, indicating its potential as a safe and effective molecular tool for cell reprogramming and cancer therapy.
BIOPROCESS AND BIOSYSTEMS ENGINEERING
(2021)
Article
Multidisciplinary Sciences
Igor Kolotilin
Summary: Plant-based production of recombinant human proteins is feasible and offers scalable, cost-effective, and eco-friendly manufacturing method for therapeutic proteins. The produced plantakines show biological functionality and can modulate pro-inflammatory cytokine secretion.
SCIENTIFIC REPORTS
(2022)
Article
Multidisciplinary Sciences
Krishna Kumar Haridhasapavalan, Pradeep Kumar Sundaravadivelu, Neha Joshi, Nayan Jyoti Das, Anshuman Mohapatra, Udayashree Voorkara, Vishwas Kaveeshwar, Rajkumar P. Thummer
Summary: This study reports the generation of recombinant human HAND2 fusion protein using a heterologous system. The purified protein retained its secondary structure and could transduce cells and translocate to the nucleus. Additionally, the recombinant protein showed angiogenic potential and synergistically activated the α-MHC promoter.
SCIENTIFIC REPORTS
(2022)
Article
Biochemistry & Molecular Biology
Hyeryeong Gye, Heeyeon Baek, Seunghyun Han, Haeun Kwon, Trang Vu Thien Nguyen, Le Thanh Mai Pham, Seunghyun Kang, Youn Hwa Nho, Dong Woog Lee, Yong Hwan Kim
Summary: Recently, there has been a growing demand for a safe and effective method of skin whitening in the cosmetics industry. Researchers have been focusing on using enzymes for melanin decolorization as an alternative to commonly used tyrosinase-inhibiting chemicals due to their low toxicity and selective decolorization ability. In this study, a recombinant lignin peroxidase (LiP) from Phanerochaete chrysosporium (PcLiP) was expressed as 10 different isozymes, and PcLiP isozyme 4 (PcLiP04) was found to have high stability and activity under conditions similar to human skin. Results showed that PcLiP04 exhibited significantly higher efficiency in melanin decolorization compared to a well-known lignin peroxidase (PcLiP01) in realistic human skin-mimicking environment. Moreover, experiments indicated that the decolorization of melanin by PcLiP04 disrupted its structure, potentially affecting its functional properties.
Article
Biology
Masamichi Nagae, Tetsuya Hirata, Hiroaki Tateno, Sushil K. Mishra, Noriyoshi Manabe, Naoko Osada, Yuko Tokoro, Yoshiki Yamaguchi, Robert J. Doerksen, Toshiyuki Shimizu, Yasuhiko Kizuka
Summary: X-ray crystallography, NMR, and computer modeling were used to investigate the structure and catalytic mechanism of GnTIVa, an enzyme involved in the synthesis of GlcNAc branches. The enzyme contains a C-terminal lectin domain that regulates its activity. The lectin domain was found to interact selectively with N-glycans having a beta 1-4 GlcNAc branch. Our findings provide insights into the biosynthesis of branching structures in N-glycans.
COMMUNICATIONS BIOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Yin Yin Siew, Amrita Rai, Han Bin Pek, Dave Siak-Wei Ow, Wei Zhang
Summary: The new and efficient purification process for recombinant human insulin production significantly improved efficiency and achieved higher recovery and purity by exploring new resins and optimizing purification steps. The process is more economical and practical than conventional methods, especially when using Nuvia HR-S cation exchanger.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Donghang Deng, Wen Li, Xiaopeng Li, Xin Yuan, Liang Li, Jiwen Wang, Chunchun Han, Shenqiang Hu
Summary: This study obtained highly pure native goose PRL and revealed a dual role of PRL in regulating the proliferation and apoptosis of goose granulosa cells, depending on its concentration and the stage of follicle development.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Review
Plant Sciences
Ryan J. Coates, Mark T. Young, Simon Scofield
Summary: Recombinant proteins play a crucial role in research, industry, and medicine. Transgenic plants have emerged as viable and preferable options for producing recombinant proteins due to their low cost, scalability, and ability to produce proteins with complex post-translational modifications. This review focuses on the recent progress in producing recombinant proteins in plants and highlights the factors that need to be considered for optimizing expression and purification of target proteins.
FRONTIERS IN PLANT SCIENCE
(2022)
Article
Multidisciplinary Sciences
Prakash G. Kshirsagar, Mansi Gulati, Wade M. Junker, Abhijit Aithal, Gaelle Spagnol, Srustidhar Das, Kavita Mallya, Shailendra K. Gautam, Sushil Kumar, Paul Sorgen, Krishan K. Pandey, Surinder K. Batra, Maneesh Jain
Summary: This study details a method for purifying biologically-active rMUC4 beta and analyzes its structure and interaction with cellular proteins. The research provides valuable tools for studying MUC4 function and generating domain-specific antibodies and vaccines for cancer immunotherapy.
SCIENTIFIC REPORTS
(2021)
Article
Multidisciplinary Sciences
Arthur Schveitzer Ferreira, Amanda Lopacinski, Michel Batista, Priscila Mazzocchi Hiraiwa, Beatriz Gomes Guimaraes, Nilson Ivo Tonin Zanchin
Summary: Epidermal growth factors (EGF) play important roles in various aspects, including embryogenesis, skin development, and immune response homeostasis. However, recombinant production of these factors in prokaryotic systems is challenging due to their structural organization. This study presents a reliable method for producing seven human growth factors of the EGF family in Escherichia coli, demonstrating their folded and stable conformation and high activity at low concentrations.
SCIENTIFIC REPORTS
(2022)
Article
Chemistry, Multidisciplinary
Junyang Gai, Esma Dervisevic, Citsabehsan Devendran, Victor J. Cadarso, Moira K. O'Bryan, Reza Nosrati, Adrian Neild
Summary: Sperm motility is an important predictor of male fertility potential, and current methods to improve it are lacking. This study successfully increased sperm motility using high frequency ultrasound, without significant effects on sperm viability and DNA integrity.
Article
Chemistry, Physical
Sushant Powar, Farin Yazdan Parast, Ashwin Nandagiri, Avinash S. Gaikwad, David L. Potter, Moira K. O'Bryan, Ranganathan Prabhakar, Julio Soria, Reza Nosrati
Summary: This study introduces an automated platform that uses thin-lens approximation and high-speed dark field microscopy to reconstruct sperm flagellar waveform in 3D, enabling analysis of sperm swimming behavior. The study found that head-tethered mouse sperm exhibit a rolling beating behavior in 3D, and discovered the properties of their flagellar waveform.
Article
Obstetrics & Gynecology
Laura Kasak, Kristiina Lillepea, Liina Nagirnaja, Kenneth Aston, Peter N. Schlegel, Joao Goncalves, Filipa Carvalho, Daniel Moreno-Mendoza, Kristian Almstrup, Michael L. Eisenberg, Keith A. Jarvi, Moira K. O'Bryan, Alexandra M. Lopes, Donald F. Conrad, Margus Punab, Maris Laan
Summary: This study analyzed the load, distribution, and clinical value of secondary findings identified in exome sequencing of patients with non-obstructive azoospermia (NOA). The findings showed that 3.6% of NOA cases carried medically actionable secondary findings, and 37% of patients with secondary findings had variants in genes linked to male infertility. Further research is needed to determine the characteristics of these secondary findings in the general population and NOA patients specifically.
HUMAN REPRODUCTION
(2022)
Article
Oncology
Amelie V. Villard, Anthony Genna, Justine Lambert, Marianna Volpert, Agnes Noel, Brett Hollier, Myriam Polette, Aline M. Vanwynsberghe, Christine Gilles
Summary: CD44 regulates the expression of tissue factor (TF) in tumor cells, promoting procoagulant activity and metastatic seeding. CD44 acts at the transcriptional level to modulate TF expression, which is important for cancer research.
Article
Immunology
Meriem Hasmim, Malina Xiao, Kris Van Moer, Akinchan Kumar, Alexandra Oniga, Michel Mittelbronn, Caroline Duhem, Anwar Chammout, Guy Berchem, Jean Paul Thiery, Marianna Volpert, Brett Hollier, Muhammad Zaeem Noman, Bassam Janji
Summary: This study reveals a new mechanism of CD73 regulation in triple-negative breast cancer, suggesting that CD73 blocking could be an attractive therapeutic strategy.
FRONTIERS IN IMMUNOLOGY
(2022)
Article
Developmental Biology
Samuel R. Cheers, Anne E. O'Connor, Travis K. Johnson, D. Jo Merriner, Moira K. O'Bryan, Jessica E. M. Dunleavy
Summary: This study reveals the crucial role of spastin, a member of the meiotic AAA ATPase protein clade, in regulating microtubule dynamics during spermatogenesis. Spastin loss leads to a complete loss of functional germ cells, enlarged spermatid nuclei with aneuploidy, and severe abnormalities in manchette structure and acrosome biogenesis. These findings have potential relevance to individuals carrying spastin variants and to the medically assisted reproductive technology industry.
Article
Public, Environmental & Occupational Health
Sarah N. Biggs, Joanne Kennedy, Sharon L. Lewis, Stephen Hearps, Moira K. O'Bryan, Robert McLachlan, Simon von Saldern, Georgina Chambers, Jane Halliday
Summary: Approximately 1 in 20 men are sub-fertile or infertile, but the causes of male infertility are not well understood. This study aims to recruit 1000 Australian male partners seeking help from fertility clinics to investigate the factors associated with male infertility. The findings will provide important information for clinicians and inform public policy on prevention and treatment strategies for infertile men.
REPRODUCTIVE HEALTH
(2023)
Article
Cell Biology
Nicola D. Kolaitis, Bethany J. Finger, D. Jo Merriner, Joseph Nguyen, Brendan J. Houston, Moira K. O'Bryan, Jessica M. Stringer, Nadeen Zerafa, Ngoc Nguyen, Karla J. Hutt, Gerard A. Tarulli, Mark P. Green
Summary: This study found that chronic exposure to an environmentally relevant concentration of the herbicide atrazine affects male fertility in mice, particularly by altering the expression of steroidogenic genes in the testicles. Further research should be conducted to investigate the multigenerational effects of atrazine on male fertility.
Article
Developmental Biology
Jessica E. M. Dunleavy, Maddison Graffeo, Kathryn Wozniak, Anne E. O'Connor, D. Jo Merriner, Joseph Nguyen, Ralf B. Schittenhelm, Brendan J. Houston, Moira K. O'Bryan
Summary: This study reveals that multiple katanin A-subunit paralogs play important roles in various processes during mammalian spermatogenesis, including the construction of the meiotic spindle, cytokinesis, midbody abscission, and spermatid remodelling. The study also identifies specific roles of certain genes in sperm flagellum development and interaction networks with other proteins.
Article
Urology & Nephrology
Sarah Kimmins, Richard A. Anderson, Christopher L. R. Barratt, Hermann M. Behre, Sarah R. Catford, Christopher J. De Jonge, Geraldine Delbes, Michael L. Eisenberg, Nicolas Garrido, Brendan J. Houston, Niels Jorgensen, Csilla Krausz, Ariane Lismer, Robert I. Mclachlan, Suks Minhas, Tim Moss, Allan Pacey, Laerke Priskorn, Stefan Schlatt, Jacquetta Trasler, Leonardo Trasande, Frank Tuettelmann, Monica Hebe Vazquez-Levin, Joris A. Veltman, Feng Zhang, Moira K. O'Bryan
Summary: Currently, there is a lack of understanding and awareness about the causes of male infertility and the importance of male reproductive health. This leads to standardized treatment strategies using medically assisted reproduction techniques, which overlook the potential co-morbidities and shift the burden to women. To improve male reproductive health, it is essential to establish centers for data and sample collection and study the effects of genetics, epigenetics, and environment on fertility and offspring health. Personalized treatment protocols should be developed and education about infertility should be increased.
NATURE REVIEWS UROLOGY
(2023)
Article
Urology & Nephrology
Margot J. Wyrwoll, Godfried W. van der Heijden, Csilla Krausz, Kenneth I. Aston, Sabine Kliesch, Robert McLachlan, Liliana Ramos, Donald F. Conrad, Moira K. O'Bryan, Joris A. Veltman, Frank Tuettelmann
Summary: Discovering the genetic causes of non-syndromic male infertility and linking them with clinical data is crucial for better understanding the condition. The International Male Infertility Genomics Consortium has developed a standardized vocabulary based on the Human Phenotype Ontology (HPO) to facilitate communication and improve the classification of male infertility. This work will contribute to the systematic recording of patients' phenotypes and the discovery of novel genetic causes for non-syndromic male infertility.
NATURE REVIEWS UROLOGY
(2023)
Article
Biology
Madeleine J. De Jong, Lesley A. Alton, Craig R. White, Moira K. O'Bryan, David G. Chapple, Bob B. M. Wong
Summary: Incubation temperature has lasting effects on growth, locomotor performance, and metabolic rate of skink offspring. Cool and hot incubation temperatures result in faster growth and larger maximum size, while hot incubation temperatures reduce locomotor performance. Effects on metabolic rate are present in sub-adults, with higher rates in cool-incubated lizards. Additionally, cool and hot incubation treatments result in shorter sperm midpieces and heads.
PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES
(2023)
Article
Chemistry, Multidisciplinary
Farin Yazdan Parast, Avinash S. Gaikwad, Ranganathan Prabhakar, Moira K. O'Bryan, Reza Nosrati
Summary: Using microfluidics, this study investigates individual sperm flagellar dynamics under the influence of flow and viscosity. The findings suggest that viscosity has a more dominant influence on flagellar waveform, and the synergistic effect of flow and viscosity promotes energy-efficient beating behavior.
CELL REPORTS PHYSICAL SCIENCE
(2023)
Article
Cell Biology
Vanessa Tsui, Ruqian Lyu, Stevan Novakovic, Jessica M. Stringer, Jessica E. M. Dunleavy, Elissah Granger, Tim Semple, Anna Leichter, Luciano G. Martelotto, D. Jo Merriner, Ruijie Liu, Lucy McNeill, Nadeen Zerafa, Eva R. Hoffmann, Moira K. O'Bryan, Karla Hutt, Andrew J. Deans, Jorg Heierhorst, Davis J. McCarthy, Wayne Crismani
Summary: Meiotic crossovers are necessary for accurate chromosome segregation and generating new allelic combinations. This study reveals the role of the tumor suppressor FANCM as an anti-crossover factor in mammals and shows that Fancm deficiency leads to an increase in crossover frequencies.
Meeting Abstract
Biochemistry & Molecular Biology
Samuel Cheers, Miguel Xavier, Manon Oud, Petra de Vries, Bilal Alobaidi, Lois Batty, Liina Nagirnaja, Don Conrad, Kenneth Aston, Corinna Friedrich, Birgit Stallmeyer, Frank Tuettelmann, Godfried van der Heijden, Moira O'Bryan, Joris Veltman
EUROPEAN JOURNAL OF HUMAN GENETICS
(2023)
