ABSTRACT
Lanthanides, a series of 15 f-block elements, are crucial in modern technology, and their purification by conventional chemical means comes at a significant environmental cost. Synthetic biology offers promising solutions. However, progress in developing synthetic biology approaches is bottlenecked because it is challenging to measure lanthanide binding with current biochemical tools. Here we introduce LanTERN, a lanthanide-responsive fluorescent protein. LanTERN was designed based on GCaMP, a genetically encoded calcium indicator that couples the ion binding of four EF hand motifs to increased GFP fluorescence. We engineered eight mutations across the parent construct's four EF hand motifs to switch specificity from calcium to lanthanides. The resulting protein, LanTERN, directly converts the binding of 10 measured lanthanides to 14-fold or greater increased fluorescence. LanTERN development opens new avenues for creating improved lanthanide-binding proteins and biosensing systems.
Subject(s)
Lanthanoid Series Elements , Lanthanoid Series Elements/metabolism , Calcium/metabolism , ProteinsABSTRACT
Human infants are born to breastfeed. While 50% of lactating persons struggle to make enough milk, there are no governmentally-approved drugs to enhance lactation1. Here, we engineer a variant of the naturally-occurring driver of lactation, the hormone Prolactin, to increase its serum half-life and produce a viable drug candidate. Our engineered variant, Prolactin-eXtra Long-acting (Prolactin-XL), is comprised of endogenously active human prolactin fused to an engineered human IgG Fc domain designed to overcome the unique drug development challenges specific to the lactating person-infant dyad. Our Prolactin-XL has a serum half-life of 70.9h in mice, 2,625-fold longer than endogenously active prolactin alone (70.9h v. 0.027h). We demonstrate that Prolactin-XL increases milk production and restores growth of pups fed by dams with pharmacologically-ablated lactation. We show that Prolactin-XL-enhanced lactation is accompanied by reversible, lactocyte-driven changes in mammary gland morphology. This work establishes long-acting prolactins as a potentially powerful pharmacologic means to combat insufficient lactation.
ABSTRACT
To grow and divide, cells must extract resources from dynamic and unpredictable environments. Many organisms use different metabolic strategies for distinct contexts. Budding yeast can produce ATP from carbon sources by mechanisms that prioritize either speed (fermentation) or yield (respiration). Withdrawing glucose from exponentially growing cells reveals variability in their ability to switch from fermentation to respiration. We observe two subpopulations of glucose-starved cells: recoverers, which rapidly adapt and resume growth, and arresters, which enter a shock state characterized by deformation of many cellular structures, including mitochondria. These states are heritable, and on high glucose, arresters grow and divide faster than recoverers. Recoverers have a fitness advantage during a carbon source shift but are less fit in a constant, high-glucose environment, and we observe natural variation in the frequency of the two states across wild yeast strains. These experiments suggest that bet hedging has evolved in budding yeast.
Subject(s)
Adaptation, Physiological , Models, Biological , Saccharomyces cerevisiae/physiology , Cell Division/physiology , Fermentation/physiology , Glucose/metabolism , Glycolysis/physiology , Metabolic Networks and Pathways/physiologySubject(s)
Disclosure , Editorial Policies , Peer Review, Research , Confidentiality , Humans , Peer Review , PublicationsABSTRACT
Experimental tools that are designed to perturb biological functions are often used to understand how cells change over time. However, in two recent papers, Segall-Shapiro et al. (2018; in Nature Biotechnology) and Rullan et al. (2018; in this issue of Molecular Cell) present tools engineered to keep cells the same. Both tools achieve perfect adaptation, that is, the ability to hold a value, in this case gene expression, constant over time despite disruptions and disturbances. Together, these papers highlight how influential dynamics can be when shaping biological functions, even when the goal is to stay constant.
Subject(s)
Gene Expression Regulation , OptogeneticsABSTRACT
A quantitative approach that tunes DNA damage strength and observes cell-cycle kinetics in single, unperturbed cells yields a new framework for thinking about cell-cycle checkpoints.