The Foundation for Food and Agriculture Research (FFAR) announced on November 30 that they are awarding $1M to a project based at the University of California, Davis, to study protein kinases of rice plants. The team is led by Dr. Pamela Ronald, a leading expert in plant genetics who has engineered disease- and flood-resistant rice. This project aims to address the growing agricultural problem of water scarcity by gaining a better understanding of the role kinases play in enabling drought-resistance. Promega will be supporting this research by providing NanoBRET™ products to help characterize kinase inhibitors.
Principal Investigator Pamela Ronald, Ph.D. Photo Credit: Deanne Fitzmaurice
The research team will begin by screening over 1,000 human kinase inhibitors to determine which ones do interact with the plant kinome and, if applicable, which kinase(s) they inhibit. Once the compound library has been established, the team will assess the inhibitors’ phenotypic effects on rice to identify kinases that, when inhibited, positively impact root growth and development. The long-term goal is to use these findings to engineer drought-resistant rice.
Four point MMOA screen for tideglusib and GW8510.
Time dependent inhibition was evaluated by preincubation of TbGSK3β with 60 nM tideglusib and 6 nM GW-8510 with 10μM and 100μM ATP. (A). Tideglusib [60 nM] in 10μM ATP. (B). GW8510 [60 nM] in 10μM ATP. (C.) Tideglusib [60 nM] at 100μM ATP. (D.) GW8510 [60 nM] at 100μM ATP. All reactions preincubated or not preincubated with TbGSK3β for 30 min at room temperature. Black = 30 min preincubation Grey = No preincubation.
The first small-molecule kinase inhibitor approved as a cancer therapeutic, imatinib mesylate (Gleevec® treatment), has been amazingly successful. However, a thorough understanding of its molecular mechanism of action (MMOA) was not truly obtained until more than ten years after the molecule had been identified.
Understanding the MMOA for a small-molecule inhibitor can play a major role in optimizing a drug’s development. The way a drug actually works–the kinetics of binding to the target molecule and how it competes with endogenous substrates of that target–ultimately determines whether or not a a candidate therapeutic can be useful in the clinic. Drugs that fail late in development are extremely costly.
Drug research and discovery for neglected tropical diseases suffer from a lack of a large commercial market to absorb the costs of late-stage drug development failures. It becomes very important to know as much as possible, simply and quickly, about MMOA for candidate molecules for these diseases that are devastating to large populations.
One such neglected topical disease is Human African trypanosomiasis (HAT, also known as sleeping sickness). Continue reading
Drug research and development is a complex and expensive process that begins with initial screening steps of candidate chemical compounds, and compounds that appear to have the desired potency against a specific cellular target or pathway are further evaluated. Candidate compounds that fail late in development or during clinical trials because of off-target effects are costly, and can be dangerous. Therefore drug developers not only need to ensure that a candidate compound is effective as a therapy, but also they need to predict any potential undesirable side effects due to off-target activities as early as possible in the drug discovery and development process. Continue reading
Image used with permission of Dr. Hongdi Liu.
Recently Promega hosted a special guest from Duke University—Dr. Neil. L. Spector, one of leading scientists in the field of breast cancer research. In a simple way
Dr. Spector presented advances in this field of cancer research
and informed us of new treatments that have the potential to improve patient lives. Continue reading
If you could design the ideal kinase assay system what would it look like?
- Would it be able to match, point for point, the results of the tried-and-true isotopic assay methods but not have any of the associated safety and waste disposal issues?
- Would it avoid the use of specific antibodies?
- Would it minimize false hits associated with many of the fluorescence-based assays?
- Would it be affordable technology, adaptable to any laboratory’s throughput from 96-well to 1,536-well automated screening?
- Would it be universal—able to assess the function of any kind of kinase (protein, lipid or sugar) that uses any kind of substrate?
- Would it be able to detect low conversion rates (low-activity enzymes) with a high signal-to-background ratio?
- Would you be able to use it with substrates that are multiphosphorylated?
If you answered “yes” to any of the above questions, you might want to take a look at the Promega Webinar “Enabling Kinase Research with a Luminescent ADP Detection Platform and Complete Kinase Enzyme Systems”, presented by Hicham Zegzouti, PhD, research scientist at Promega. Here he describes the ADP-Glo™ Assay platform, which meets these and several other criteria.
The precise molecular lesion that occurs with the Philadelphia Chromosome translocation—a rearrangement that creates a bcr-abl fusion in which the abl tyrosine kinase is constitutively active leading to the development of chronic myeloid leukemia is the first description of dysregulation of a kinase leading to a particular disease state. However, the human genome contains 518 protein kinases and many other atypical kinases, and one-third of all human proteins are phosphorylated. It is now estimated that over 400 human diseases are caused by dysregulation or mutation of kinases, making kinases a major target for drug discovery efforts. Continue reading