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This book presents modern and classic analytical approaches that are crucial for the biochemical and functional characterization of the archetypal protein, glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The distinguishing feature of the book is that it covers, in addition to other methods, some of the uncommon but valuable techniques as well. For example, in-gel visualization of enzyme activity, immunoblotting protocols for native (non-denatured) proteins, and proteins resolved by pH-gradient [IEF-isoelectrofocusing], etc. These expedient methods are relevant and vital for the verification of biochemical properties of GAPDH, or similar protein of interest. This work outlines detailed protocols that are essential to investigate classical (cellular) and recently reported extracellular (secretory) isoforms of GAPDH. Precisely, the book covers techniques pertinent to enzymatic and non-enzymatic analysis of GAPDH that include, but not limited to, electrophoretic mobility shift assay (EMSA), two-dimensional (2D)-immunoblotting, immunofluorescence/confocal microscopy, mass spectrometry, ion-exchange and affinity chromatography. Readers will discover the importance of the experimental methods described in the book as they relate to the evaluation of the role and significance of GAPDH. Furthermore, majority of the methods described in the book have also been validated in the author’s laboratory, besides other research groups worldwide, underlining the repeatability and reproducibility of the protocols. Each method begins with an abstract and a brief background emphasizing its application and relevance. This will enable the readers to determine the choice of experimental design according to their research objectives. The book explains the methods systematically with ample illustrations to facilitate quick and easy comprehension of the practical knowledge. Although the book is focused on GAPDH, many of the protocols may be adopted to other proteins or enzymes with minimal modifications. Noteworthy, it is unequivocally established that GAPDH is a multifunctional protein involved in several cellular processes of health & disease conditions. Hence, this book will be a valuable practical guide for young researchers, scientists and clinician-scientists.
The book represents a comprehensive review and synthesis of the biomedical literature that spans over a half-century on a single protein called glyceraldehyde 3-phosphate dehydrogenase (or, GAPDH). Due to the protein’s involvement in a vast array of cellular activities, GAPDH is of interest to the cell biologist, immunologist, virologist, biochemist etc. The protein has a significant role in fertility, cancer and neurodegeneration, suggesting that this book can be a vital resource for drug development. GAPDH function may provide insight into anesthesia. Furthermore, GAPDH is highly conserved meaning that the protein found in microorganisms, such as pathogens, remained relatively unchanged in evolution. Pathogens use GAPDH as a virulence factor, offering a unique challenge in developing anti-microbial agents that target this protein. To the evolutionary biologist, a book on the multi-functionality of GAPDH provides a focal point for a cogent discussion on the very origin of life.
Malignant melanoma is the most aggressive type of skin cancer with invasive growth patterns. Recent advances in melanoma management have focused on cell growth, proliferation, migration, and survival biomarkers. Numerous FDA-approved molecular-targeted therapies, such as tyrosine kinase inhibitors (TKIs), have been developed targeting genetic biomarkers that play a critical role in tumorigenesis when overexpressed. The use of targeted therapies as an alternative or adjuvant to immunotherapy has revolutionized the management of metastatic melanoma. Although this treatment strategy is more efficacious and less toxic than traditional therapies, targeted therapies are less effective after prolonged treatment due to acquired resistance given the mutations and activation of resistant mechanisms in melanoma tumors. In this sense, identifying novel target therapies and repositioned drugs that could modulate the disease progression, the potential molecular and pharmacological mechanisms of therapies are of utmost interest.
Crop plants are constantly exposed to multiple abiotic (such as drought, salinity, cold, flooding, heavy metal, and heat) and/or biotic (bacterial/fungal/viral) stress factors that hinder their growth and development, subsequently leading to decreases in quality and yield. During the last two decades, many classical genetic and breeding approaches have been used to develop stress-tolerant and climate-adaptable plants that can provide a better yield to meet food demands. Climate change poses a major risk to food security as the world faces frequent floods, droughts, heat waves, and the emergence of new invasive pests and diseases. Novel genomic and genetic approaches look promising to improve plant resilience under stress conditions and achieve sustainable crop improvements. Recent advances in sequencing technologies have facilitated the generation of a plethora of genomic resources in a variety of crop and plant species. With the increased availability of genomic and transcriptomic data, an increasing number of quantitative trait loci and candidate genes are being identified for their application in improving plant tolerance to abiotic and biotic stresses. New approaches such as genomic selection and genomic-assisted breeding have been utilized to develop stress-tolerant cultivars in a variety of plant species. Furthermore, transgenics and rapidly evolving CRISPR technology offer great potential for plant improvement. This Research Topic aims to provide insights into the molecular and genetic factors involved in imparting abiotic and biotic stress tolerance in plants and their application in enhancing plant adaptation to these stress conditions. To review the progress in this research category, we invite manuscripts related to the plant responses to abiotic/biotic stresses and trait improvement through genomic selection, and transgenic or gene-editing approaches. Studies including physiological, biochemical, and molecular genetic analyses revealing the mechanisms involved in plant response to abiotic/biotic stresses are welcome. Topic editor Dr. Balaji Aravindhan Pandian is employed by Enko Chem Inc. All other Topic Editors declare no competing interests with regard to the Research Topic subject.
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